Anti-endotoxin compounds

ABSTRACT

Disclosed are lipid A analogs useful for the treatment of septic shock and LPS-mediated activation of viral infection.

This application is a divisional of Ser. No. 07/935,050, filed 25 Aug.1992, now U.S. Pat. No. 5,530,113, which is a continuation in part ofSer. No. 07/776,100 filed 11 Oct. 1991, now abandoned.

BACKGROUND OF THE INVENTION

The invention relates to compounds which are useful as anti-endotoxicdrugs, particularly analogs of lipid A.

The incidence of gram negative bacteremia in the United States has beenestimated to be approximately 100,000 to 300,000 cases per year, with amortality rate of 30-60% (Dudley, Am. J. Hosp. Pharm. 47, Supp.3:S3,1990). Antibiotics are commonly used as the primary chemotherapy forthis disease; however, their bactericidal action results in disruptionof the bacterium and concomitant release of endotoxin, i.e., thelipopolysaccharide (LPS) moiety of the bacterial outer membrane. Theliberated LPS induces a number of pathophysiological events in mammals(collectively referred to as gram-negative endotoxemia or sepsissyndrome): these include fever, generalized inflammation, disseminatedintravascular coagulation (DIC), hypotension, acute renal failure, acuterespiratory distress syndrome (ARDS), hepatocellular destruction, andcardiac failure (Dudley, supra; Braunwald et al., eds., Harrison'sPrinciples of Internal Medicine, 11th ed., McGraw-Hill Book Co., NewYork, 1987).

Although the endotoxin initiates sepsis, it has little or no directeffect on tissues; instead, it triggers a cascade of biologic mediatorswhich lead to sepsis and septic shock. Endotoxin stimulates monocytesand macrophages to produce tumor-necrosis factor and interleukin-1, twomajor primary mediators. These mediators then cause the sepsis syndromeby stimulating inflammatory or other cells, such as endothelial cells,to secrete a cascade of secondary mediators (e.g., prostaglandins,leukotrienes, interferons, platelet-activating factor, endorphins, andcolony-stimulating factors). These inflammatory mediators influencevasomotor tone, microvascular permeability, and the aggregation ofleukocytes and platelets. Although the actions and interactions of thesesubstances appear to be complex, their net effect in initiating septicshock appears to be very significant (Braunwald et al., supra).

As reported by DiPiro (Am. J. Hosp. Pharm. 47, Supp.3:S6, 1990), thebacterial lipopolysaccharide molecule has three main regions: along-chain polysaccharide (O Antigen) region, a core region, and a lipidA region. The entire lipopolysaccharide molecule and some of itscomponents have toxic effects. Most of these toxic effects, however, arebelieved to be attributable to the lipid A portion. Structurally, lipidA is composed of a disaccharide and acylated by long-chain fatty acids.

Therapies for endotoxin-related diseases have generally been directedtoward controlling the inflammatory response. Such therapies include:corticosteroid treatment, suggested to ameliorate endotoxin-mediatedcell membrane injury and to reduce production of certain biologicmediators (Bone,N, Eng. J. Med. 317:653, 1987; Veterans AdministrationSystemic Sepsis Cooperative Study Group, N. Eng. J. Med. 317:659, 1987;Braunwald et al., supra); administration of antibodies designed toneutralize the bacterial LPS endotoxin (see, e.g., Ziegler et al., N.Eng. J. Med. 307:1225, 1982); treatment with naloxone, which apparentlyblocks the hypotensive effects associated with the sepsis syndrome(Sheagren et al., Shock Syndromes Related to Sepsis. In: Wyngaarden andSmith, eds., Cecil Testbook of Medicine, 18th ed. Philadelphia, 1988, pp1538-41); and treatment with nonsteroidal anti-inflammatory drugs,purported to block cyclo-oxygenases and thereby decrease the productionof certain secondary mediators such as prostaglandins and thromboxane(DiPiro, supra).

SUMMARY OF THE INVENTION

In general, the invention features a compound of the formula: ##STR1##wherein at least one R¹, R², R³, or R⁴ is: ##STR2##

wherein each L is O, N, or C; each M is O or N; each E, independently,is an integer between 0 and 14 inclusive; each G, independently, is N,O, S, SO, or SO₂ ; each m, independently, is an integer between 0 and 14inclusive; each n, independently, is an integer between 0 and 14inclusive; each p, independently, is an integer between 0 and 10inclusive; and each q, independently, is an integer between 0 and 10inclusive;

each of the remaining R¹, R², R³, and R⁴, independently, is ##STR3##

wherein each L is O, N, or C; each M is O or N; each x, independently,is an integer between 0 and 14 inclusive; each y, independently, is aninteger between 0 and 14 inclusive; each z, independently, is an integerbetween 0 and 10 inclusive; and each G, independently, is N, O, S, SO,or SO₂ ;

each A¹ and A², independently, is H, OH, OCH₃, ##STR4##

wherein each d, independently, is an integer between 0 and 5 inclusive;each f, independently, is an integer between 0 and 5 inclusive; each g,independently, is an integer between 0 and 5 inclusive; and each A³,independently, is: ##STR5##

(CH₂)_(j) --CO₂ H, or O--(CH₂)_(j) --CO₂ H

wherein each j, independently, is an integer between 0 and 14 inclusive;

X is H, (CH₂)_(t) CH₃, (CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃, (CH₂)_(t)OPO (OH)₂,

(CH₂)_(t) --CH═CH--(CH₂)_(v) CH₃, (CH₂)_(t) --O--R⁵, ##STR6##

wherein each t and v, independently, is an integer between 0 and 14inclusive; and R⁵ is any of the possibilities listed above for R¹ --R⁴ ;and

Y is H, OH, O(CH₂)_(w) CH₃, a halo group ##STR7##

wherein w is an integer between 0 and 14 inclusive.

Preferably, at least one R¹, R², R³, or R⁴ is: ##STR8##

wherein each m, independently, is an integer between 0 and 10 inclusive;each n, independently, is an integer between 0 and 10 inclusive; and foreach p and q, independently, 0≦(p+q)≦12;

each of the remaining R¹, R², R³, and R⁴, independently, is: ##STR9##

wherein each x, independently, is an integer between 0 and 10 inclusive;each z, independently, is an integer between 0 and 3 inclusive; and eachG, independently, is SO or SO₂ ;

each A¹ and A², independently, is: ##STR10##

wherein each d, independently, is an integer between 0 and 2 inclusive;

X is H, (CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃ or (CH₂)_(t) CH₃, whereint is an integer between 0 and 6 inclusive and v is an integer between 0and 6; and

Y is a halo group or OH.

More preferably, at least one R¹, R², R³, or R⁴ is: ##STR11##

wherein each n, independently, is an integer between 6 and 10 inclusive,most preferably 6; and 6≦(p+q)≦10, and most preferably, q is 5;

each of the remaining R¹, R², R³, and R⁴, independently, is ##STR12##

wherein each x, independently, is an integer between 6 and 11 inclusive,most preferably 6 or 10; and each G, independently, is SO or SO₂ ;

each A¹ and A², independently, is: ##STR13##

X is CH₂ OH, CH₂ OCH₃, or CH₂ O(CH₂)_(v) CH₃, wherein v is an integerbetween 1 and 3 inclusive; and

Y is OH.

Most preferably, the above compounds are formulated as a lysine salt, ATris salt, an ammonium salt, or a sodium salt; and include lipid Aanalogs B274, B276, B286, B288, B313, B314, B379, B385, B387, B388,B398, B400, B479, B214, B218, B231, B235, B272, B287, B294, B300, B318,B377, B380, B406, B410, B425, B426, B427, B442, B451, B452, B459, B460,B464, B465, B466, B531, B415, B718, B587, B737, B736, B725, and B763(described herein).

Of the compounds of the first aspect (above), those which may beisolated from natural sources (e.g., from Rhodopseudomonas capsulata orRhodopseudomonas sphaeroides) are less preferred in the invention.

In a second aspect, the invention features a compound of the formula:##STR14##

wherein at least one R¹, R², R³, or R⁴ is: ##STR15##

wherein each L is O, N, or C; each M is O or N; each m, independently,is an integer between 0 and 14 inclusive; each n, independently, is aninteger between 0 and 14 inclusive; each p, independently, is an integerbetween 0 and 10 inclusive; and each q, independently, is an integerbetween 0 and 10 inclusive; each x, independently, is an integer between0 and 14; each y, independently, is an integer between 0 and 14inclusive; and each z, independently, is an integer between 0 and 10inclusive;

each of the remaining R¹, R², R³, and R⁴, independently, is: ##STR16##

wherein each L is O, N, or C; each M is O or N; each E, independently,is an integer between 0 and 14 inclusive; each m, independently, is aninteger between 0 and 14 inclusive; each n, independently, is an integerbetween 0 and 14 inclusive; each p, independently, is an integer between0 and 10 inclusive; and each q, independently, is an integer between 0and 10 inclusive; each x, independently, is an integer between 0 and 14inclusive; each y, independently, is an integer between 0 and 14inclusive; each z, independently, is an integer between 0 and 10inclusive; and each G, independently, is N, O, S, SO, or SO₂ ;

each A¹ and A², independently, is H, OH, ##STR17##

wherein each d, independently, is an integer between 0 and 5 inclusive;each f, independently, is an integer between 0 and 5 inclusive; each g,independently, is an integer between 0 and 5 inclusive; and each A³,independently, is; ##STR18##

(CH₂)_(j) --CO₂ H, or O--(CH₂)_(j) --CO₂ H

wherein each j, independently, is an integer between 0 and 14 inclusive;

X is H, (CH₂)_(t) CH₃, (CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃,

(CH₂)_(t) --CH═CH--(CH₂)_(v) CH₃, (CH₂)_(t) --O--R⁵, ##STR19##

wherein each t and v, independently, is an integer between 0 and 14inclusive; and R⁵ is any of the possibilities listed above for R¹ -R⁴ ;and

Y is H, OH, O(CH₂)_(w) CH₃, a halo group, ##STR20##

wherein w is an integer between 0 and 14 inclusive.

Preferably, at least one R¹, R², R³, or R⁴ is: ##STR21##

wherein each m, independently, is an integer between 0 and 10 inclusive;each n, independently, is an integer between 0 and 10 inclusive, mostpreferably 6; and for each p and q, independently, 0≦(p+q)≦12, and mostpreferably, q is 5;

each of the remaining R¹, R², R³, and R⁴, independently, is: ##STR22##

wherein each x, independently, is an integer between 0 and 10 inclusive,most preferably 6 or 10; and each z, independently, is an integerbetween 0 and 3 inclusive;

each A¹ and A², independently, is: ##STR23##

wherein each d, independently, is an integer between 0 and 2 inclusive;

X is H, (CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃ or (CH₂)_(t) CH₃, whereint is an integer between 0 and 6 inclusive and v is an integer between 0and 6; and

Y is OH.

Most preferably, the compounds of the second aspect are formulated as alysine salt, a Tris salt, an ammonium salt, or a sodium salt; andinclude lipid A analogs: B415, B459, B460, B465, B466, B477, B479, B510,B427, B464, and B531 (described herein).

In a third aspect, the invention features a therapeutic compositionwhich includes, as an active ingredient, a compound according to to theinvention formulated in a physiologically-acceptable carrier.

In a fourth aspect, the invention features a compound of formula:##STR24## wherein R² is: ##STR25##

wherein each J, independently, is OH or a protected OH; each L is O, N,or C; each M is O or N; each E, independently, is an integer between 0and 14 inclusive; each m, independently, is an integer between 0 and 14inclusive; each n, independently, is an integer between 0 and 14inclusive; each p, independently, is an integer between 0 and 10inclusive; each q, independently, is an integer between 0 and 10inclusive; each x, independently, is an integer between 0 and 14inclusive; each y, independently, is an integer between 0 and 14inclusive; each z, independently, is an integer between 0 and 10inclusive; and each G, independently, is N, O, S, SO, or SO₂ ;

P¹ is CH, a protected OH, or a protected A¹ group, wherein A¹ is:##STR26##

wherein each d, independently, is an integer between 0 and 5 inclusive;each f, independently, is an integer between 0 and 5 inclusive; each g,independently, is an integer between 0 and 5 inclusive; and each A³,independently, is: ##STR27##

(CH₂)_(j) --CO₂ H, or O--(CH₂)₂ --CO₂ H

wherein each j, independently, is an integer between 0 and 14 inclusive;and

P² is H, a halo group, OH, a protected OH, O(CH₂)_(w) CH₃, ##STR28##

wherein w is an integer between 0 and 14 inclusive.

Preferably, R² is: ##STR29##

wherein each J, independently, is OH or a protected OH; each m,independently, is an integer between 0 and 10 inclusive; each n,independently, is an integer between 0 and 10 inclusive; each x,independently, is an integer between 0 and 10 inclusive; each z,independently, is an integer between 0 and 3 inclusive; each G,independently, is SO or SO₂ ; and for each p and q, independently,O≦(p+q)≦12.

P¹ is OH, a protected OH, or a protected A¹ group, wherein A¹ is:##STR30##

wherein each d, independently, is an integer between 0 and 2 inclusive;and

P² is H, OH, a protected OH, or O(CH₂)_(w) CH₃, wherein w is an integerbetween 0 and 3 inclusive.

Most preferably, R² is ##STR31##

wherein each J, independently, is OH or a protected OH; each x,independently, is an integer between 6 and 11 inclusive; and each G,independently, is SO or SO₂ ; each n, independently, is an integerbetween 6 and 10 inclusive; and 6≦(p+q)≦10;

P¹ is OH, a protected OH, or a protected A¹ group, wherein A¹ is:##STR32##

p² is H, OH, a protected OH, or OCH₃.

In a fifth aspect, the invention features a compound of the formula:##STR33## wherein R⁴ is: ##STR34##

wherein each J, independently, is OH or a protected OH; each L is O, N,or C; each M is O or N; each E, independently, is an integer between 0and 14 inclusive; each m, independently, is an integer between 0 and 14inclusive; each n, independently, is an integer between 0 and 14inclusive; each p, independently, is an integer between 0 and 10inclusive; each q, independently, is an integer between 0 and 10inclusive; each x, independently, is an integer between 0 and 14inclusive; each y, independently, is an integer between 0 and 14inclusive; each z, independently, is an integer between 0 and 10inclusive; and each G, independently, is N, O, S, SO, or SO₂ ;

P³ is OH, a protected OH, OCH₃, an A² ' group, or a protected A² 'group, wherein A² ' is: ##STR35##

wherein each d, independently, is an integer between 0 and 5 inclusive;each f, independently, is an integer between 0 and 5 inclusive; each g,independently, is an integer between 0 and 5 inclusive; and each A³,independently, is: ##STR36##

(CH₂)_(j) --CO₂ H, or O--(CH₂)_(j) --CO₂ H

wherein each j, independently, is an integer between 0 and 14 inclusive;and

Z is OH, a protected OH, an activated OH, or a displaceable leavinggroup; and

X' is X or a protected X group, wherein the X group is H, (CH₂)_(t) CH₃,(CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃, (CH₂)_(t) OPO(OH)₂,

(CH₂)_(t) --CH═CH--(CH₂)_(v) CH₃, (CH₂)_(t) --O--R⁵, ##STR37##

wherein each t and v, independently, is an integer between 0 and 14inclusive; and R⁵ is any of the possibilities listed above for R¹ -R⁴.

Preferably, R⁴ is: ##STR38##

wherein each J, independently, is OH or a protected OH; each m,independently, is an integer between 0 and 10 inclusive; each n,independently, is an integer between 0 and 10 inclusive; each x,independently, is an integer between 0 and 10 inclusive; each z,independently, is an integer between 0 and 3 inclusive; each G,independently, is SO or SO₂ ; and for each p and q, independently,0≦(p+q)≦12;

P³ is H, OH, a protected OH, an A² ' group, or a protected A² ' group,wherein A² ' is: ##STR39##

wherein each d, independently, is an integer between 0 and 2 inclusive;and

X' is H, (CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃ or (CH₂)_(t) CH₃,wherein t is an integer between 0 and 6 inclusive and v is an integerbetween 0 and 6.

Most preferably, R⁴ is: ##STR40##

wherein each J, independently, is OH or a protected OH; each x,independently, is an integer between 6 and 11 inclusive; and each G,independently, is SO or SO₂ ; each n, independently, is an integerbetween 6 and 10 inclusive; and 6≦(p+q)≦10;

P³ is OH, a protected OH, an A² ' group, or a protected A² ' group,wherein A² ' is: ##STR41##

X' is CH₂ OH, CH₂ OCH₃, or CH₂ O(CH₂)_(v) CH₃, wherein v is an integerbetween 1 and 3 inclusive.

In a sixth aspect, the invention features a compound of the formula:##STR42## wherein each R² and R⁴, independently is: ##STR43##

wherein each J, independently, is OH or a protected OH; each L is O, N,or C; each M is O or N; each E, independently, is an integer between 0and 14 inclusive; each m, independently, is an integer between 0 and 14inclusive; each n, independently, is an integer between 0 and 14inclusive; each p, independently, is an integer between 0 and 10inclusive; each q, independently, is an integer between 0 and 10inclusive; each x, independently, is an integer between 0 and 14inclusive; each y, independently, is an integer between 0 and 14inclusive; each z, independently, is an integer between 0 and 10inclusive; and each G, independently, is N, O, S, SO, or SO₂ ;

P¹ is OH, a protected OH, or a protected A¹ group; and

P³ is OH, a protected OH, an A² ' group, or a protected A² ' group,wherein each A¹ and A² ' group, independently, is: ##STR44##

wherein each d, independently, is an integer between 0 and 5 inclusive;each f, independently, is an integer between 0 and 5 inclusive; each g,independently, is an integer between 0 and 5 inclusive; and each A³,independently, is: ##STR45##

(CH₂)_(j) --CO₂ H, or O--(CH₂)_(j) --CO₂ H

wherein each j, independently, is an integer between 0 and 14 inclusive;and

P² is H, a halo group, OH, a protected OH, O(CH₂)_(w) CH₃, ##STR46##

wherein w is an integer between 0 and 14 inclusive;

Q, independently, is N₃ or NH₂ ; and

X' is X or a protected X group, wherein the X group is H, (CH₂)_(t) CH₃,(CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃,

(CH₂)_(t) --CH═CH--(CH₂)_(v) CH₃, (CH₂)_(t) --O--R⁵, ##STR47##

wherein each t and v, independently, is an integer between 0 and 14inclusive; and R⁵ is any of the possibilities listed above for R¹ -R⁴.

Preferably, each R² and R⁴, independently, is: ##STR48##

wherein each J, independently, is OH or a protected OH; each m,independently, is an integer between 0 and 10 inclusive; each n,independently, is an integer between 0 and 10 inclusive; each x,independently, is an integer between 0 and 10 inclusive; each z,independently, is an integer between 0 and 3 inclusive; each G,independently, is SO or SO₂ ; and for each p and q, independently,0≦(p+q)≦12;

P¹ is OH, a protected OH, or a protected A¹ group; and

P³ is OH, a protected OH, an A² ' group, or a protected A² ' group,wherein each A¹ and A² ' group, independently, is: ##STR49##

wherein each d, independently, is an integer between 0 and 2 inclusive;

P² is H, OH, a protected OH, or O(CH₂)_(w) CH₃, wherein w is an integerbetween 0 and 3 inclusive; and

X' is H, (CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃ or (CH₂)_(t) CH₃,wherein t is an integer between 0 and 6 inclusive and v is an integerbetween 0 and 6.

Most preferably, each R² and R⁴, independently, is ##STR50##

wherein each J, independently, is OH or a protected OH; each x,independently, is an integer between 6 and 11 inclusive; and each G,independently, is SO or SO₂ ; each n, independently, is an integerbetween 6 and 10 inclusive; and 6≦(p+q)≦10;

P¹ is OH, a protected OH, or a protected A¹ group; and

P³ is OH, a protected OH, an A² ' group, or a protected A² ' group,wherein each A¹ and A² ' group, independently, is: ##STR51##

P² is OH; and

X' is CH₂ OH, CH₂ OCH₃, or CH₂ O(CH₂)_(v) CH₃, wherein v is an integerbetween 1 and 3 inclusive.

In a seventh aspect, the invention features a compound of the formula:##STR52## wherein each R¹, R², R³, and R⁴, independently is: ##STR53##

wherein each J, independently, is OH or a protected OH; each L is O, N,or C; each M is O or N; each E, independently, is an integer between 0and 14 inclusive; each m, independently, is an integer between 0 and 14inclusive; each n, independently, is an integer between 0 and 14inclusive; each p, independently, is an integer between 0 and 10inclusive; each q, independently, is an integer between 0 and 10inclusive; each x, independently, is an integer between 0 and 14inclusive; each y, independently, is an integer between 0 and 14inclusive; each z, independently, is an integer between 0 and 10inclusive; and each G, independently, is N, O, S, SO, or SO₂ ;

P¹ is OH, a protected OH, or a protected A¹ group; and

P³ is OH, a protected OH, an A₂ ' group, or a protected A² ' group,wherein each A¹ and A² ' group, independently, is: ##STR54##

wherein each d, independently, is an integer between 0 and 5 inclusive;each f, independently, is an integer between 0 and 5 inclusive; each g,independently, is an integer between 0 and 5 inclusive; and each A³,independently, is: ##STR55##

(CH₂)_(j) --CO₂ H, or O--(CH₂)_(j) --CO₂ H

wherein each j, independently, is an integer between 0 and 14 inclusive;and

P² is H, a halo group, OH, a protected OH, O(CH₂)_(w) CH₃, ##STR56##

wherein w is an integer between 0 and 14 inclusive; and

X' is X or a protected X group, wherein the X group is H, (CH₂)_(t) CH₃,(CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃,

(CH₂)--CH═CH--(CH₂)_(v) CH₃, (CH₂)_(t) --O--R⁵, ##STR57##

wherein each t and v, independently, is an integer between 0 and 14inclusive; and R⁵ is any of the possibilities listed above for R¹ -R⁴.##STR58##

wherein each J, independently, is OH or a protected OH; each m,independently, is an integer between 0 and 10 inclusive; each n,independently, is an integer between 0 and 10 inclusive; each x,independently, is an integer between 0 and 10 inclusive; each z,independently, is an integer between 0 and 3 inclusive; each G,independently, is SO or SO₂ ; and for each p and q, independently,0≦(p+q)≦12;

P¹ is OH, a protected OH, or a protected A¹ group; and

P³ is OH, a protected OH, an A² ' group, or a protected A² ' group,wherein each A¹ and A² ' group, independently, is: ##STR59##

wherein each d, independently, is an integer between 0 and 2 inclusive;

P² is H, OH, a protected OH, or O(CH₂)_(w) CH₃, wherein w is an integerbetween 0 and 3 inclusive; and

X' is H, (CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃ or (CH₂)_(t) CH₃,wherein t is an integer between 0 and 6 inclusive and v is an integerbetween 0 and 6.

Most preferably, each R¹, R², R³, and R⁴, independently, is ##STR60##

wherein each J, independently, is OH or a protected OH; each x,independently, is an integer between 6 and 11 inclusive; and each G,independently, is SO or SO₂ ; each n, independently, is an integerbetween 6 and 10 inclusive; and 6≦(p+q)≦10;

P¹ is OH, a protected OH, or a protected A¹ group; and

P³ is OH, a protected OH, an A² ' group, or a protected A² ' group,wherein each A¹ and A² ' group, independently, is: ##STR61##

P² is OH; and

X' is CH₂ OH, CH₂ OCH₃, or CH₂ O(CH₂)_(v) CH₃, wherein v is an integerbetween 1 and 3 inclusive.

In an eighth aspect, the invention features a method of making acompound of the formula ##STR62## involving the steps of (a) providing amannopyranoside of the formula ##STR63## and

(b) reacting said mannopyranoside with a catalytic amount of napthalenein the presence of lithium.

In an ninth aspect, the invention features a method of making a compoundof the formula ##STR64## involving the steps of (a) providing a compoundof the formula ##STR65## and

(b) reacting the compound with ammonium cerium nitrate and an azidealkali metal salt, preferably, sodium azide.

In a preferred embodiment, the method further involves the step ofreacting ##STR66## with sodium nitrate to form ##STR67##

In a tenth aspect, the invention features a method of selectively makingthe α-stereoisomer of the compound of formula ##STR68## involving thesteps of: (a) providing a compound of formula ##STR69## (b) dissolvingthe compound in trichloracetonitrile; and (c) reacting the dissolvedcompound with lithium bis(trimethylsilyl)amide.

In an eleventh aspect, the invention features a method of coupling a3,4-dimethoxybenzyl protecting group so an activated azido saccharideinvolving reacting the azido saccharide first with dimethoxybenzylalcohol and then with boron trifluoride etherate. In a preferredembodiment, the azido saccharide is ##STR70##

In a twelfth aspect, the invention features a method of coupling anallyloxycarbonate protecting group to a hydroxyl sidechain of asaccharide involving reacting the saccharide first with phosgene andthen with allyl alcohol.

in preferred embodiments, the saccharide is an azido saccharide; thesaccharide is of the formula ##STR71## the saccharide is of the formula##STR72## and the saccharide is of the formula ##STR73##

In a thirteenth aspect, the invention features a method of selectivelyremoving a t-butyldimethylsilyl protecting group from an acyl-protectedsaccharide involving reacting the saccharide with hydrofluoric acid.

In preferred embodiments, the saccharide is a disaccharide; the acylprotecting group is an allyloxycarbonate group; the acyl-protectedsaccharide is ##STR74## the acyl-protected saccharide is ##STR75## theacyl-protected saccharide is ##STR76## the acyl-protected saccharide is##STR77## the acyl-protected saccharide is ##STR78## the acyl-protectedsaccharide further includes a 3,4-dimethoxybenzyl protecting group; andthe acyl-protected saccharide is ##STR79##

In a fourteenth aspect, the invention features a method of coupling abis(alkoxy)phosphonyl sidechain to a saccharide involving reacting thesaccharide first with a bis(alkoxy) (diisopropylamino)phosphine andtetrazole and then with an oxidant.

In preferred embodiments, the bis(alkoxy)phosphonyl sidechain is anallyloxy-protected phosphate group; the oxidant is m-chloroperoxybenzoicacid; the saccharide is a disaccharide, preferably, an azido saccharide;the azido saccharide is of the formula ##STR80## the azido saccharide isof the formula ##STR81## the azido saccharide is of the formula##STR82## and the azido saccharide is of the formula ##STR83##

In a fifteenth aspect, the invention features a method of removing a3,4-dimethoxybenzyl protecting group from an azido saccharide involvingreacting the azido saccharide with2,3-dichloro-5,6-dicyano-1,4-benzoquinone in the dark under anaerobicconditions.

In a sixteenth aspect, the invention features a method of removing a3,4-dimethoxybenzyl protecting group from an azido saccharide involvingreacting the azido saccharide with ammonium cerium nitrate.

In a preferred embodiment of the fifteenth and sixteenth aspects, theazido saccharide is ##STR84##

In a seventeenth aspect, the invention features a method for selectivelycoupling an α-trichloroimidate activating group to an azido sugarinvolving reacting the azido sugar with trichloroacetonitrile and cesiumcarbonate. In a preferred embodiment, the azido sugar is of the formula##STR85##

In an eighteenth aspect, the invention features a method for making adisaccharide involving the steps: (a) providing an azido monosaccharidehaving a 3,4-dimethoxybenzyl protecting group and a free hydroxyl group;and (b) reacting the 3,4-dimethoxybenzyl-protected azido monosaccharidewith a second activated azido monosaccharide under an argon atmospherein the presence of boron trifluoride etherate or trimethylsilyltrifluoromethanesulfonate.

In preferred embodiments, the azido monosaccharides are ##STR86## andthe azido monosaccharides are ##STR87##

In a nineteenth aspect, the invention features a method for reducing anazido sidechain of a saccharide without reducing an unsaturatedsidechain involving reacting the azido saccharide with atin(II)tris-arylthiolate trialkylamine complex in the dark underanaerobic conditions.

In preferred embodiments, the tin(II)tris-benzenethiolate trialkylamineis tin(II)tris-benzenethiolate triethylamine; the azido saccharide is adisaccharide; the disaccharide is of the formula ##STR88## thedisaccharide is of the formula ##STR89## and the disaccharide is of theformula ##STR90##

In a twentieth aspect, the invention features a method for removing anallyloxy protecting group from a saccharide molecule involving thesteps: (a) providing a saccharide having an allyloxy-protected hydroxylgroup; and (b) reacting the protected saccharide with a palladiumcomplex.

in preferred embodiments, the palladium complex istetrakis(triphenylphosphine)palladium(O); the saccharide is of theformula ##STR91## and the saccharide is of the formula ##STR92##

In a twenty-first aspect, the invention features a method for alkylatingthe C₆ hydroxyl of a hexose without alkylating other free hydroxylgroups involving reacting the hexose with a silver salt and an alkylhalide.

In preferred embodiments, the silver salt is silver oxide or silvercarbonate; the alkyl halide is methyl iodide; and the hexose is of theformula ##STR93##

In a twenty-second aspect, the invention features a method ofphosphorylating the C₁ carbon of a saccharide having an amido sidechainincluding a β-sulfoxy group involving reacting the amido saccharidefirst with a lithium base under anaerobic conditions in the cold andthen with dialkyl chlorophosphate.

In preferred embodiments, the lithium base is lithiumbis(trimethylsilyl)amide; the dialkyl chlorophosphate is diallylchlorophosphate; and the saccharide is of the formula ##STR94##

In a twenty-third aspect, the invention features a method of making a C₁dialkylphosphonate saccharide involving (a) first reacting thesaccharide with trichloroacetonitrile and carbonate under anaerobicconditions; and then (b) treating with a Lewis acid and atrialkylphosphite under anaerobic conditions.

In preferred embodiments, the carbonate is cesium carbonate; thetrialkylphosphite is triallylphospshite; and the saccharide is of theformula ##STR95##

IN a twenty-fourth aspect, the invention features a method for couplingan alkyl sidechain to an azido saccharide having a free hydroxylinvolving reacting the azido saccharide with an alkali metal salt and asulfonyl mono-activated alkyl diol under anaerobic conditions.

In preferred embodiments, the alkali metal salt is sodium hydride; andthe sulfonyl mono-activated alkyl diol is monotosyl alkyl diol.

In a twenty-fifth aspect, the invention features a method for treating adisease in a mammal for which a lipid A receptor antagonist is effectiveinvolving administering to the mammal a therapeutic composition of theinvention in a dosage effective to reduce the binding of LPS to a lipidA receptor.

In a twenty-sixth aspect, the invention features a method for treatingseptic shock in a mammal involving administering to the mammal atherapeutic composition of the invention in a dosage effective toantagonize LPS-mediated target cell activation.

In a twenty-seventh aspect, the invention features a method for treatingLPS-mediated activation of a viral infection in a mammal involvingadministering to the mammal a therapeutic composition of the inventionin a dosage effective to antagonize LPS-mediated target cell activation.

In preferred embodiments, the virus includes an NF-KB binding site in areplication control sequence; the virus is a human immunodeficiencyvirus, for example, HIV-1 or HIV-2; the virus is a herpes virus, forexample, a Herpes simplex virus; and the virus is an influenza virus.

A "protected" group, as used herein, means a group (e.g., a hydroxylgroup attached to an intermediate compound of the invention) which isprevented from undergoing a chemical reaction; the particular reactionwhich is blocked and the conditions under which the protecting group isremoved are particular to each intermediate compound and are madeobvious to those skilled in the art by the synthetic proceduresdescribed herein. Examples of preferred protecting groups include, butare not limited to, methyl, benzyl, substituted benzyl, silyl,alkylsilyl, methoxymethyl, alkylacyl, alky oxy carbonyl, and aromaticacyl groups.

By "activated" is meant having a carbon center which is adjacent to a"displaceable leaving group". The choice of an appropriate leaving groupis made obvious to one skilled in the art by the synthetic proceduresdescribed herein. Examples of preferred leaving groups include, but arenot limited to, acyl oxy, iminoether oxy, iminocarbonate oxy, phenoxy,sulfonyl oxy, alkyl thio, and aryl thio, aryl oxy, Se alkyl, and halogroups.

By "mono activated" is meant a compound (e.g., an intermediate compoundof the invention) which has only one activated group (as defined above)attached.

The lipid A analogs described herein provide particularly potenttherapeutics for the treatment or prevention of LPS-mediated disorders.Without being held to any particular theory, the analogs likely act byblocking access to LPS target sites on mediator molecules, therebycompeting directly with bacterial LPS. Because this block occurs at avery early step in the mediator cascade, the therapy is unusuallyeffective and is accompanied by few or no side effects. In addition,because the lipid A analogs of the instant invention are synthesizedchemically, they are relatively inexpensive to manufacture and are ofunusually high purity and defined chemical constitution, resulting inlow immunoreactivity.

Other features and advantages of the invention will be apparent from thefollowing description of the preferred embodiments and from the claims.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

There now follow examples of synthetic compounds which are particularlyuseful in the invention. These examples are designed to illustrate, notlimit, the invention.

Table 1 provides abbreviations which are used throughout thespecification.

                  TABLE 1                                                         ______________________________________                                        Ac             Acetate                                                                        ##STR96##                                                     Sph            thiophenyl                                                                     ##STR97##                                                     DMB            3,4-dimethoxybenzyl                                                            ##STR98##                                                     AOC            allyloxycarbonate                                                              ##STR99##                                                     TBS            t-butyldimethylsilyl                                                           ##STR100##                                                    AllylO         allyloxy                                                                       ##STR101##                                                    MPM            p-methoxybenzyl                                                                ##STR102##                                                    ______________________________________                                    

EXAMPLE 1

This example illustrates the synthesis of the lipid A analogs describedherein as well as the synthesis of novel product intermediates, alsoclaimed in the invention.

PART A Preparation of Monosaccharides and Disaccharides ##STR103##

To a 0° C. solution of D-mannose (i.e., Compound 1; 1.5 kg, 8.33 mol;Lancaster Chemical Co., Windham, N.H.) dissolved in anhydrous pyridine(3.5 kg, 126.0 mol, Aldrich Chemical Co., Milwaukee, Wis.) was added 5.6kg (54.9 mol) acetic anhydride (Aldrich Chemical Co.), over four hoursat a rate to maintain the reaction temperature between 20°-40° C. Theresulting solution was then stirred at room temperature overnight; 5.0 g(40.9 mmol) of 4-dimethylaminopyridine (Aldrich Chemical Co.) was added;the resulting mixture was stirred for an additional 48 hours. Thereaction mixture was then poured into 14.0 L of ice-water under vigorousstirring, and extracted with 12.0 L dichloromethane (J. T. Baker, Inc.,Phillipsburg, N.J.). The organic layer was washed first with 1Nhydrochloric acid (10.0 L; Fisher Scientific Co., Pittsburgh, Pa.), thenwith saturated aqueous sodium bicarbonate solution (20.0 L; FisherScientific Co.), and finally with 5.0 L saturated aqueous sodiumchloride solution and dried over 3 kg sodium sulfate (Fisher ScientificCo.). The solution was then filtered through a glass fritted funnel andconcentrated under reduced pressure at 40° C. to provide 3.5 kg ofCompound 2 {R_(f) : 0.39 [ethyl acetate (J. T. Baker, Inc.): hexanes (J.T. Baker, Inc), 1:1(v/v)]} as a brown oil which was utilized in the nextstep without purification. ##STR104##

Compound 2 (3.0 kg, 7.3 mol) was mixed with thiophenol (Aldrich ChemicalCo.) (1.5 L, 11.0 mol), dissolved in 8.0 L of chloroform (J. T. Baker,Inc.), and boron trifluoride etherate (1.6 L, 12.9 mol, Aldrich ChemicalCo.) was added at such a rate that the reaction temperature remainedbelow 40° C. Upon complete consumption of the starting material [asmeasured by thin layer chromatographic analysis using hexanes:ethylacetate, 1:1(v/v)], the mixture was cooled to room temperature andslowly poured, with rapid mechanical stirring, into 15.0 L of saturatedaqueous sodium bicarbonate solution. The resulting mixture was extractedwith 18.0 L of dichloromethane, and the organic layer washed first with15.0 L of saturated aqueous sodium bicarbonate solution and then 10.0 Lof saturated aqueous sodium chloride solution, and the resultantsolution dried over 3 kg sodium sulfate and filtered through a glassfritted funnel. The filtrate was concentrated under reduced pressure at40° C. to provide 4.99 kg of Compound 3 (R_(f) : 0.63 [ethylacetate:hexanes, 1:1(v/v)]; as a dark brown oil which was utilized inthe next step without further purification. ##STR105##

To a mechanically stirred solution of Compound 3 (6.28 kg, 4.3 moldissolved in methyl alcohol 13.0 L; Aldrich Chemical Co.) was graduallyadded 750.0 mL (3.28 mol) of a 25%(wt/v) sodium methoxide/methyl alcoholsolution (Aldrich Chemical Co.), maintaining a reaction temperaturebelow 40° C. The resulting mixture was stirred at 40° C. untilcompletion, i.e., until only material having an R_(f) of 0.05 (by thinlayer chromatography analysis using ethyl acetate) was detected. Themixture was then cooled to room temperature and neutralized by additionof Dowex acidic 50X 8-200 ion exchange resin (Aldrich Chemical Co.). Theneutralized reaction mixture was filtered through a glass fritted funneland then concentrated under reduced pressure at 40° C. to yield a brownoil. The oil was partially purified by the addition, with stirring, oftwo 10.0 aliquots of 5:1(v/v ethyl acetate/hexanes; the supernatant wasdiscarded after each wash. The product, Compound 4 {R_(f) : 0.05 [ethylacetate]}, was obtained as a brown oil after drying under vacuum.##STR106##

Compound 4 (3.0 kg) was dissolved in anhydrous N,N-dimethylformamide(6.0 L; Aldrich Chemical Co.) at room temperature. To this solution wasfirst added 1.0 kg (4.3 mol) (±)-10-camphorsulfonic acid (AldrichChemical Co.), and next added (slowly, over 48 hours) 8.0 L of2,2-dimethoxypropane (Aldrich Chemical Co.). Thin layer chromatographicanalysis [ethyl acetate:hexanes, 1:4(v/v)] indicated completion of thereaction. The completed reaction mixture was poured into 10.0 L ofsaturated aqueous sodium bicarbonate solution and extracted with 12.0 Ldichloromethane. The organic layer was washed first with 5.0 L water andthen with 10.0 L of saturated aqueous sodium chloride solution, anddried over 3.0 kg sodium sulfate. The dried solution was filteredthrough a glass fritted funnel and concentrated under reduced pressureat 40° C. to provide Compound 5 as a black oil. The crude oil wasdissolved in 10.0 L of boiling ethyl acetate, cooled to roomtemperature, and allowed to crystallize overnight. The crystalline masswas cooled to 5° C., filtered, and washed with 5.0 L of hexanes at 0° C.to provide 2.0 kg of partially purified Compound 5 as light brownneedles. The remaining filtrate was concentrated under reduced pressureat 40° C., the resultant oil dissolved in 2.0 dichloromethane, and thesolution applied to a short pad of silica gel (2.0 kg; J. T. Baker,Inc.) and eluted with 1:4(v/v) ethyl acetate/hexanes. The filtrate wasconcentrated and crystallized from ethyl acetate, yielding an additional1.5 kg of crystalline product. The combined crystals were recrystallizedfrom ethyl acetate to provide a total of 2.8 kg of Compound 5 {R_(f):0.60 [ethyl acetate:hexanes, 1.4(v/v)]} in 66% overall yield fromCompound 1. ##STR107##

Compound 5 (1.98 kg, 6.0 mol) was dissolved in anhydrous tetrahydrofuran(6.0 L; Aldrich Chemical Co.), and 40.0 g (0.31 mol) of naphthalene(Aldrich Chemical Co.) was added at room temperature under a nitrogenatmosphere. To the solution was next added 20.0 g (2.9 mol) of lithiumwire (3.2 mm diameter, 0.01% sodium; Aldrich Chemical Co.), cut into 20cm long pieces, and the resulting mixture was subjected to rapidmechanical stirring. Upon completion of the reaction {as monitored bythin layer chromatographic analysis [ethyl acetate:hexanes, 1:1(v/v)]},excess lithium wire was removed, and the reaction mixture was pouredinto 10.0 L of saturated aqueous ammonium chloride solution (FisherScientific Co.). The mixture was then extracted with 10.0 L ofdichloromethane; the organic layer was washed with 7.0 L of saturatedaqueous sodium chloride solution, dried over 2.0 kg sodium sulfate,filtered through a fritted glass funnel, and concentrated under reducedpressure at room temperature to provide 1.4 kg of crude Compound 6{R_(f) : 0.50 [ethyl acetate:hexanes, 1:1(v/v)]}. ##STR108##

Compound 6 (3.5 kg) was slowly added to a mechanically stirred mixtureof 0.5 L acetic anhydride and 4.5 L anhydrous pyridine. The addition wascarried out in an ice-water bath in order to maintain a reactiontemperature under 25° C. Forty-eight hours later the reaction mixturewas concentrated to dryness under reduced pressure at room temperatureto yield a syrupy, crystalline mass, which was filtered on a frittedglass funnel and washed with 1.0 L hexanes (at 0°) to provide 2.25 kg ofCompound 7 as white needles {R_(f) : 0.5 [ethyl acetate:hexanes,1:4(v/v)]}. ##STR109##

Compound 7 (50.6 g, 0.22 mol) was dissolved in 1.3 L anhydrousacetonitrile (Aldrich Chemical Co.), and a mixture of finely powderedammonium cerium nitrate (550.0 g, 1.0 mol, Aldrich Chemical Co.) andsodium azide (40.0 g, 0.62 mol, Aldrich Chemical Co.) was added at -30°C., using a solid additional funnel. During the addition, the reactiontemperature rose to -26° C. After stirring at -28° C. for four hours,the reaction mixture was poured slowly into 4.0 L ice water. Evolutionof gas was observed during this process. The mixture was then dilutedwith 4.0 L ethyl acetate, and the two layers were separated. The organiclayer was washed first with a 1.0 L portion of water, then with 2.0saturated aqueous sodium bicarbonate solution, and finally with 1.0 Lsaturated aqueous sodium chloride solution; the resulting solution wasthen dried over 500.0 g sodium sulfate, filtered through a fritted glassfunnel, and concentrated to dryness under reduced pressure, at roomtemperature to provide approximately 70.0 g of crude product as a lightyellow oil. The oil was passed through a short pad of silica gel (1.0kg) with a mixture of 1:2(v/v) ethyl acetate:hexanes. Evaporation ofsolvent from the product-containing fractions (as identified by thinlayer chromatographic analysis) under reduced pressure at roomtemperature and drying overnight under vacuum at room temperatureprovided 56.0 g (0.169 mol) of (Compound 8) as a colorless foam in 77%yield. ##STR110##

compound 8 (56.0 g, 0.168 mol) was dissolved in a mixture of dioxane(1.47 L; Aldrich Chemical Co.) and water (600.0 mL), and 64.5 g (0.93mol) sodium nitrite (Aldrich Chemical Co.) was added. The reactionmixture was refluxed for one hour, cooled to room temperature, anddiluted with ethyl acetate (2.0 L). The two layers were separated, andthe aqueous layer extracted with 2.0 L ethyl acetate. The combinedorganic layers were washed first with 1.0 L water, then with 1.0 Lsaturated aqueous sodium bicarbonate solution, and finally with 1.0 Lsaturated aqueous sodium chloride solution; the solution was dried over500.0 g sodium sulfate, filtered through a fritted glass funnel, andconcentrated under reduced pressure at room temperature to yield ayellow oil. The oil was passed through a short pad of silica gel (1.0kg) with a mixture of 1:1 (v/v) ethyl acetate/hexanes. Evaporation ofsolvent from the product-containing fractions (as identified by thinlayer chromatographic analysis) under reduced pressure at roomtemperature and drying overnight under vacuum at room temperatureprovided 55.0 g (0.168 mol) of Compound 9 {R_(f) : 0.14 [ethylacetate:hexanes, 1:4(v/v)]} as a colorless foam in a near quantitativeyield. ##STR111##

Compound 9 (1.50 g, 5.20 mmol) was dissolved in anhydroustetrahydrofuran (20.0 mL) and trichloroacetonitrile (14 ml, 0.14 mol;Aldrich Chemical Co.). To this solution was added 1.8 mL (1.8 mmol) of a1.0M solution of lithium bis(trimethylsilyl)amide (Aldrich Chemical Co.)in hexanes added at 0° C. over four hours. The reaction was quenchedwith 10.0 mL saturated aqueous ammonium chloride solution and extractedwith 200.0 mL ethyl acetate. The organic layer was washed with 100.0 mLsaturated aqueous sodium chloride solution, dried over 50.0 g sodiumsulfate, filtered, and concentrated under reduced pressure at roomtemperature. The crude product was purified on a silica gel (150.0 g)column, eluting with 1:3(v/v) ethyl acetate/hexanes to provide 1.40 g(3.2 mmol) of the α-trichloroimidate, i.e., Compound 10a {R_(f) : 0.37[hexanes:ethyl acetate, 3:1 (v/v)]}, as a syrup, in 67% yield, and 0.47g (1.09 mmol) of the β-trichloroimidate, i.e., Compound 10b {R_(f) :0.45 [hexanes:ethyl acetate, 3:1 (v/v)]}, as crystalline needles, in 25%yield. ##STR112##

Compound 10a (130.0 mg, 0.30 mmol) was mixed with 3,4-dimethoxybenzylalcohol (65.0 mL, 0.45 mmol; Aldrich Chemical Co.) and anhydrousdichloromethane (5.0 mL). To this mixture was added 200.0 mg of finelypowdered AW-300 molecular sieves (Aldrich Chemical Co.). The mixture wasstirred for one hour at room temperature, cooled to -78° C., and 1.0 mLof a 0.02M dichloromethane solution of boron trifluoride etherate addedover a period of six hours. The reaction was quenched with 1.0 Lsaturated aqueous sodium bicarbonate solution and extracted with 50.0 mLdichloromethane. The organic layers were dried over 25.0 g sodiumsulfate, filtered through a fritted glass funnel, and concentrated underreduced pressure at room temperature. Purification on a silica gelcolumn by eluting with 2:1(v/v) hexanes/ethyl acetate provided a 6:1mixture of Compound 11b {R_(f) : 0.28 [hexanes:ethyl acetate, 3:1 v/v)]}and Compound 11a {R_(f) : 0.31 [hexanes:ethyl acetate, 3:1 (v/v)]} as acrystalline solid. The solid was recrystallized from 2:1(v/v)hexanes/ethyl acetate as described above. Evaporation of solvent fromthe product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 91.9 mg (0.21mmol) of pure Compound 11b in 70% yield. ##STR113##

To a solution of Compound 11b (18.48 g, 0.04 mol) in methyl alcohol(200.0 ml) was added 2.0 mL of a 25%(w/v) sodium methoxide/methylalcohol solution; the resulting mixture was stirred at room temperaturefor four hours. The reaction mixture was neutralized with 10.0 mLsaturated aqueous ammonium chloride solution and extracted with 500.0 mLethyl acetate. The organic layer was washed first with 100.0 mL water,and then with 100.0 mL saturated aqueous sodium chloride solution, anddried over 50.0 g sodium sulfate. Filtration through a cotton plug andevaporation of the solvent under reduced pressure at room temperatureyielded a crude product which was applied to a silica gel (2.0 kg)column and eluted with 2:1(v/v) hexanes/ethyl acetate to provide 15.1 g(0.038 mol) of Compound 12 {R_(f) : 0.19 [hexanes:ethyl acetate,2:1(v/v)]} in a 90% yield. ##STR114##

Compound 12 (15.1 g, 0.038 mol) was dissolved in anhydrousdichloromethane (200.0 mL). To this solution were sequentially added12.4 g (0.038 mol) of Compound A6 (see below), 9.5 g (0.046 mol)1,3-dicyclohexylcarbodiimide (Aldrich Chemical Co.), and 50.0 mg (0.41mmol) of 4-dimethylaminopyridine, at 0° C., with magnetic stirring.After 30 minutes, the mixture was diluted with 100.0 mL hexanes andfiltered through 100.0 g Celite 545 (Aldrich Chemical Co.). The filtratewas evaporated under reduced pressure at room temperature and theresidue purified on a silica gel (2.0 kg) column by elution with1:4(v/v) ethyl acetate/hexanes. Evaporation of solvent from theproduct-containing fractions (as determined by thin layerchromatographic analysis) under reduced pressure at room temperatureprovided 22.1 g (0.034 mol) Compound 13 {R_(f) : 0.41 [ethylacetate:hexanes, 1:2(v/v)]} in an 89% yield. ##STR115##

Compound 13 (22.0 g, 0.034 mol) was dissolved in glacial acetic acid(90.0 mL; Fisher Scientific Co.) and water (10.0 mL), and magneticallystirred at room temperature for 36 hours. The mixture was thenevaporated under reduced pressure at room temperature and azeotropedthree times with 50.0 mL portions of toluene (J. T. Baker, Inc.). Theresidue was purified on a silica gel (2.0 kg) column by elution with alinear gradient of 1:99(v/v) to 5:95(v/v) methyl alcohol/chloroform toprovide 22.7 g (0.037 mol) of Compound 14 {R_(f) : 0.15[chloroform:methyl alcohol, 98:2(v/v)]} in quantitative yield. Compound14 was used for the next step without further purification. ##STR116##

Compound 14 (20.6 g, 0.034 mol) was dissolved in N,N-dimethylformamide(33.9 mL), under a nitrogen atmosphere, at 0° C. To this solution wasadded 11.5 g (0.17 mol) of imidazole (Aldrich Chemical Co.) followed by5.5 g (0.037 mol) of tert-butyldimethylsilyl chloride (LithcoCorporation of America, Gastonia, N.C.). The resulting mixture wasstirred for one hour, diluted with 500.0 mL ethyl acetate, and pouredinto 500.0 mL saturated aqueous sodium bicarbonate solution. The organiclayer was washed first with 200.0 mL saturated aqueous sodiumbicarbonate solution, then with 200.0 mL water, and finally with 100.0mL saturated aqueous sodium chloride solution. The organic layer wasdried over 200.0 g sodium sulfate, filtered through a cotton plug, andevaporated under reduced pressure at room temperature. The residue wasthen purified by silica gel (2.0 kg) column chromatography, eluting with1:4 (v/v) ethyl acetate/hexanes. Evaporation of solvent from theproduct-containing fractions (as determined by thin layerchromatographic analysis) provided 24.2 g (0.033 mol) of Compound 15{R_(f) : 0.76 [ethyl acetate:hexanes, 1:1(v/v)]} in 98% yield.##STR117##

Compound 15 (24.1 g, 0.033 mol) was dissolved in anhydrous toluene(300.0.0 mL) and anhydrous pyridine (30.0 mL), at 0° C., under anitrogen atmosphere. To this solution was slowly added (i.e., over 10min) 24.1 mL (0.046 mol) of a 1.93M solution of phosgene in toluene(Fluka Chemical Corp., Ronkonkoma, N.Y.). Thirty minutes later, 24.0 mL(0.353 mol) of allyl alcohol (Fluka Chemical Corp., Ronkonkoma, N.Y.)was added over a five-minute period, and the resulting reaction mixturewas stirred for an additional 10 minutes. The reaction was quenched byaddition of 100.0 mL saturated aqueous sodium bicarbonate solution, anddiluted with 1.0 L ethyl acetate. The organic layer was washed firstwith 500.0 mL water, then with 500.0 mL saturated aqueous sodiumchloride solution, dried over 500.0 g sodium sulfate, filtered through acotton plug, and then evaporated under reduced pressure at roomtemperature. The residue was purified by silica gel (2.0 kg) columnchromatography, eluting with 1:4 (v/v) ethyl acetate/hexanes to provide25.3 g (0.031 mol) of Compound 16 {R_(f) : 0.60 [ethyl acetate:hexanes,1:2(v/v)]} in 94% yield. ##STR118##

In a 250.0 mL polypropylene tube, 25.3 g (0.031 mol) of Compound 16 wasdissolved in 100.0 mL of acetonitrile (Aldrich Chemical Co.). To thesolution, at room temperature with magnetic stirring, was added 100.0 mLof a 4M solution of hydrofluoric acid (Aldrich Chemical Co.) inacetonitrile. After 30 minutes, the reaction was quenched with 100.0 mLsaturated aqueous sodium bicarbonate solution and extracted with 500.0mL chloroform. The organic layer was washed with 100.0 mL water followedby 100.0 mL saturated aqueous sodium chloride solution, and then driedover 100.0 g sodium sulfate, filtered through a cotton plug, and thesolvent evaporated sunder reduced pressure at room temperature. Theresidue obtained was purified by silica gel (2.0 kg) columnchromatograph, eluting with 2:3(v/v) ethyl acetate/hexanes to provide19.9 g (0.029 mol) of Compound 17 {R_(f) : 0.53 [ethyl acetate:hexanes,1:1 (v/v)]} in 91% yield. ##STR119##

Compound 12 (20.0 g, 50.1 mmol) was dissolved in anhydrousdichloromethane (500.0 mL) at 0° C., and 19.4 g (52.7 mmol) of CompoundB6 (see below), 20.8 g (100.9 mmol) of 1,3-dicyclohexycarbodiimide, and120.0 mg (0.98 mmol) of 4-dimethylaminopyridine were added. Afterstirring for 20 minutes at room temperature the reaction mixture wasdiluted with 500.0 mL hexanes, filtered through 100.0 g Celite 545, andthe solids washed with 100.0 mL hexanes. The combined filtrates werethen concentrated under reduced pressure at room temperature, and theresidue obtained was purified by silica gel (2.0 kg) columnchromatography, eluting with 1:3(v/v) ethyl acetate/hexanes. Evaporationof solvent from the product containing fractions (as determined by thinlayer chromatographic analysis) under reduced pressure at roomtemperature provided 35.0 g (47.0 mmol) of Compound 18 {R_(f) : 0.54[ethyl acetate:hexanes, 1:4 (v/v)]} in 93% yield. ##STR120##

A solution of 35.0 g (47.0 mmol) of Compound 18 in a mixture of 240.0 mLof glacial acetic acid and 60.0 mL of water was magnetically stirred for14 hours at room temperature. The reaction mixture was then concentratedunder reduced pressure at room temperature, and the crude productazeotroped with three 50.0 mL portions of toluene. The product waspurified by silica gel (3.0 kg) column chromatography, eluting firstwith 1:1(v/v) hexanes/diethyl ether (Mallinckrodt Chemical Co., St.Louis, Mo.) followed by elution with ethyl acetate to provide 29.3 g(41.6 mmol) of Compound 19 {R_(f) : 0.62 [dichloromethane:methylalcohol, 95:5(v/v)]} in an 89% yield. ##STR121##

Compound 19 (4.9 g, 6.94 mmol) was dissolved in anhydrous toluene (50.0mL) and anhydrous pyridine (12.0 mL) at 0° C., under a nitrogenatmosphere. To this solution was added 1.31 mL (12.3 mmol)allylchloroformate (Aldrich Chemical Co.). After seven and a half hours,the mixture was diluted with 100.0 mL ethyl acetate and washed, firstwith 100.0 mL saturated aqueous sodium bicarbonate solution, then with100.0 mL water, and finally with 100.0 mL saturated aqueous sodiumchloride solution. The mixture was then dried over 50.0 g sodiumsulfate, and the solvents evaporated under reduced pressure at roomtemperature. The residue was dissolved in 10.0 mL dichloromethane,loaded onto a silica gel (500.0 g) column, and eluted with 1:2(v/v)ethyl acetate:hexanes. Evaporation of solvent from theproduct-containing fractions (as determined by thin layer columnchromatographic analysis) under reduced pressure at room temperatureprovided 4.2 g (5.32 mmol) of Compound 20 {R_(f) : 0.70 [ethylacetate:hexanes, 1:1(v/v)]} in 77% yield. ##STR122##

To a magnetically stirred solution of 18.26 g (0.02 mol) of Compound 20in 200.0 mL anhydrous tetrahydrofuran, 17.02 mL (0.069 mol)bis(allyloxy) (diisopropylamino) phosphine (prepared by the method ofBrannwarth and Kung, Tetrahedron Lett, 30, 4219, 1989) and 14.58 g(0.208 mol) 1H-tetrazole (Amresco Chemical Co., Solon, Ohio) were addedat room temperature under a nitrogen atmosphere. After one hour, themixture was cooled to -78° C., and a solution of 11.95 g3-chloroperoxybenzoic acid (Aldrich Chemical Co.) in 80.0 mL anhydrousdichloromethane was added. The reaction temperature was adjusted to 0°C., and the mixture stirred for 20 minutes. The reaction was quenchedwith 50.0 mL of a 10% aqueous sodium thiosulfate solution; and following10 minutes of stirring at room temperature, the mixture was warmed toroom temperature. The mixture was then poured into 200.0 mL saturatedaqueous sodium bicarbonate solution and extracted with 500.0 mLdichloromethane. The organic layer was washed first with 100.0 mL water,and then with 100.0 mL saturated aqueous sodium chloride solution, driedover 200.0 g sodium sulfate, and the solvents evaporated under reducedpressure at room temperature. The residue was purified on a silica gel(2.0 kg) column, eluting with ethyl acetate: hexanes [1:2(v/v)].Evaporation of solvent from the product-containing fractions (asdetermined by thin layer chromatographic analysis) under reducedpressure at room temperature provided 17.64 g (0.0186 mol) of Compound21 {R_(f) : 0.32 [ethyl acetate:hexanes, 1.2(v/v)]} in an 80.5% yield.##STR123##

Compound 21 (35.0 g, 0.0369 mol) was dissolved in 40.0 mLtert-butylalcohol (Aldrich Chemical Co.), 40.0 mL pH 7.0 phosphatebuffer concentrate (Fisher Scientific Co.) and 200.0 mL dichloromethane.To this solution, 33 g (0.145 mol)2,3-dichloro-5,6-dicyano-1,4-benzoquinone (Aldrich Chemical Co.) wasadded at room temperature under a nitrogen atmosphere. The mixture wasstirred in the dark at room temperature for 14 hours. The reaction wasquenched with 200.0 mL 10% sodium thiosulfate solution (FisherScientific Co.), poured into 100.0 mL saturated aqueous sodiumbicarbonate solution, and extracted with 1.0 L chloroform. The organiclayer was washed first with 200.0 mL water and then with 200.0 mLsaturated aqueous sodium chloride solution, dried over 500.0 g sodiumsulfate, and the solvents evaporated under reduced pressure at roomtemperatures. The mixture was purified on a silica gel (3.0 kg) column,eluting with 98.2(v/v) dichloromethane/methyl alcohol. Evaporation ofsolvent from the product-containing fractions (as determined by thinlayer chromatographic analysis) under reduced pressure at roomtemperature provided 27.5 g (0.0344 mol) of Compound 22 {R_(f) : 0.57[dichloromethane:methyl alcohol, 95.5(v/v)]} in a 94% yield. ##STR124##

To a mechanically-stirred solution of 52.8 g (0.066 mol) of Compound 22in 1.32 L trichloroacetonitrile, 53.0 g (0.163 mol) of cesium carbonate(Aldrich Chemical Co.) was added at room temperature under a nitrogenatmosphere. After eight hours, the mixture was filtered through 100.0 gCelite 545, washed with 500.0 mL dichloromethane, and the solventevaporated under reduced pressure at room temperature. The residue waspurified on a silica gel (3.0 kg) column eluted with 95:5(v/v)dichloromethane/diethyl ether. Evaporation of solvent from theproduct-containing fractions (as determined by thin layerchromatographic analysis) under educed pressure at room temperatureprovided 30.0 g (0.03 mol) of Compound 23A (α-isomer) {R_(f) : 0.79[dichloromethane:diethyl ether, 9:1(v/v)]} and 10.0 g (0.01 mol) ofCompound 23B (β-isomer) {R_(f) : 0.76 [dichloromethane:diethyl ether,9:1(v/v)]}. Compound 23B was subjected again to the above reaction andsubsequent purification to yield a second crop of Compound 23A. The twocrops were combined to provide a total of 32.2 g (0.034 mol) of pureCompound 23A in 52% yield. ##STR125##

A mixture of 6.0 g (6.3 mmol) of Compound 23A and 4.5 g of Compound 17(6.5 mmol) were dried under vacuum for 14 hours and dissolved in 100.0mL of anhydrous dichloromethane. To this solution was added 10.0 g ofpowdered AW-300 molecular sieves (which had been flame-dried undervacuum), and the resulting mixture magnetically stirred for one hour atroom temperature, under argon. The mixture was cooled to -23° C., and9.45 mL (1.89 mmol) of a 0.2M boron trifluoride etherate:dichloromethane solution [prepared by dissolving 0.25 mL (2.03 mmol ofboron trifluoride etherate in 10.0 mL of anhydrous dichloromethane andstirring with 200.0 mg powdered AW-300 molecular sieves for one hour atroom temperature] was added using a syringe pump over a six-hour period.The reaction was quenched with 30.0 mL of saturated aqueous sodiumbicarbonate solution, diluted with 500.0 mL dichloromethane, andfiltered through 50.0 g Celite 545. The filtrate was washed first with200.0 mL saturated aqueous sodium bicarbonate solution, then with 200.0mL water, and finally with 200.0 mL saturated aqueous sodium chloridesolution; the filtrate was then dried over 300.0 g sodium sulfate,filtered through a cotton plug, and the solvents evaporated underreduced pressure at room temperature. The resultant residue was purifiedon a silica gel (1.0 kg) column by elution with 1:3(v/v) ethylacetate/hexanes. Evaporation of solvent from the product-containingfractions (as determined by thin layer chromatographic analysis) underreduced pressure at room temperature provided 5.42 g (3.67 mmol) ofCompound 24 {R_(f) : 0.34 [ethyl acetate:hexanes, 1:2(v/v)]} in 59%yield. ##STR126##

Compound 24 (2.11 g, 1.43 mmol) was dissolved in anhydrousdichloromethane (22.0 mL), and 1.9 g of tin(II)tris-benzenethiolatetriethylamine complex (prepared by the method of Barta et al.,Tetrahedron Lett. 47, 5941, 1987) was added. The resultant mixture wasstirred for eight hours at room temperature under a nitrogen atmospherein the absence of light. Thin layer chromatographic analysis[hexanes:ethyl acetate, 1:1(v/v)] demonstrated that all startingmaterial was consumed. The reaction mixture was loaded directly onto asilica gel (200.0 g) column and eluted first with 4:1(v/v) hexanes/ethylacetate to remove reagent by-products, and then with 200.0 mL ethylacetate. Evaporation of solvent from the product-containing fractions(as determined by thin layer chromatographic analysis) under reducedpressure at room temperature provided 0.91 g (1.34 mmol) of Compound 25{R_(f) : 0.34 [ethyl acetate:hexanes, 1:1v/v)]} in 93.5% yield.##STR127##

Compound 25 (1.91 g, 1.34 mmol) was dissolved in 10.0 mL anhydrousdichloromethane. To this mixture was added 1.7 g (4.65 mmol) of CompoundC6 (see below) and 1.98 g (9.60 mmol) of 1,3-dicyclohexylcarbodiimide atroom temperature. Fourteen hours later, thin layer chromatographicanalysis [hexanes:ethyl acetate, 1:1(v/v)] indicated that the reactionwas complete. The reaction mixture was diluted with 50.0 mL ethylacetate and filtered through 10.0 g Celite 545, the solids washed with20.0 mL ethyl acetate, and the filtrate evaporated under reducedpressure at room temperature, yielding a syrupy residue. The syrup wasdissolved in 5.0 mL dichloromethane, loaded onto a silica gel (100.0 g)column, and eluted, initially with 1:2(v/v) ethyl acetate/hexanes andthen with 1:1(v/v) ethyl acetate/hexanes. Evaporation of solvent fromthe product-containing fractions (as determined by thin layerchromatographic analysis) under reduced pressure at room temperatureprovided 2.0 g (0.95 mmol) of Compound 26 {R_(f) : 0.5 [ethylacetate:hexanes, 1:1(v/v)]} in 71% yield. ##STR128##

Compound 26 (817.0 mg, 0.386 mmol) was mixed with 13.5 mL ofdichloromethane, 1.4 mL of tert-butyl alcohol and 1.4 mL of pH 7.0phosphate buffer concentrate. To the mixture was added 439.0 mg (1.9mmol) of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone. The resultingmixture was magnetically stirred under a nitrogen atmosphere in the darkuntil thin layer chromatographic analysis [dichloromethane:methylalcohol, 95:5(v/v)] indicated the complete consumption of startingmaterial (approximately four and a half hours). The reaction was thenquenched with 10.0 mL 10% aqueous sodium thiosulfate solution, dilutedwith 100.0 mL dichloromethane, and poured into 50.0 mL saturated aqueoussodium bicarbonate solution. The organic layer was separated, washedwith 50.0 mL saturated aqueous sodium chloride solution, dried oversodium sulfate, and filtered through a cotton plug. The crude reactionmixture was loaded directly onto a silica gel (100.0 g) column andeluted with 95:5(v/v) dichloromethane/methyl alcohol to provide 606.0 mg(0.35 mmol) of Compound 27 {R_(f) : 0.42 [dichloromethane:methylalcohol, 95:5(v/v)]} in 91% yield.

To produce Lipid A Analog B274-34, Compound 27 was deprotected generallyas described below for the preparation of Compound 31, and the free acidproduct was reacted with L-lysine as described below for analog B214-32.##STR129##

To a solution of 408.4 mg (0.237 mmol) of Compound 27 in anhydroustetrahydrofuran (5.0 mL), 0.265 mL (0.262 mmol) of 0.99M n-butyllithium(Aldrich Chemical Co.) in hexanes was added slowly under a nitrogenatmosphere, at -78° C., with stirring. After five minutes, 0.71 mL(0.355 mmol) of 0.5M diallyl chlorophosphate (prepared by the method ofHayakawa et al., Tetrahedron Lett. 28, 2259, 1987) in anhydrous toluenewas added, and the mixture was stirred for 10 min. The mixture was thenwarmed to 0° C., stirred 15 minutes, and the reaction quenched with 0.1mL of glacial acetic acid. The reaction mixture was poured into a 20.0mL saturated aqueous sodium bicarbonate solution and extracted with 100mL dichloromethane. The organic layer was washed with 20 mL saturatedaqueous sodium chloride solution, dried over 50.0 g sodium sulfate,filtered through a cotton plug, and the solvents evaporated underreduced pressure at room temperature. The residue obtained was purifiedon a silica gel (100.0 g) column by elution with 1:1(v/v)dichloromethane/ethyl acetate, to provide 298.2 mg (0.158 mmol) ofCompound 28 {R_(f) : 0.38 [dichloromethane:methyl alcohol, 95:5(v/v)]}in 66.8% yield.

To produce Lipid A Analogs B231-31 and B231-32 Compound 28 wasdeprotected generally as described below for the preparation of Compound31. Lipid A Analog B231-32 was produced by reacting the free acidproduct with L-lysine as described below for analog B214-32. Lipid AAnalog B231-31 was produced by reacting the free acid product with Trisas described below for B214-31. ##STR130##

389.0 mg (0.92 mmol)1,1,1-tris(acetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (prepared bythe method of Dess and Martin, J. Org. Chem. 48:4156, 1983) wasdissolved in 9.1 mL of anhydrous dichloromethane, and 600.0 mg offlame-dried powdered 4Å molecular sieves (Aldrich Chemical Co.) wereadded. To this reaction mixture was then slowly added a solution of287.2 mg (0.15 mmol) of Compound 28 dissolved in 2.9 mL dichloromethane,at 0° C., under argon. Two hours later, 5.0 mL (0.50 mmol) of a 0.1Mdichloromethane solution of1,1,1-tris(acetoxy)-1,1-dihydro-1,2-benzidoxol-3(1H)-one was slowlyadded, and the reaction mixture was stirred for an additional two hours.The reaction mixture was then diluted with 10.0 mL diethyl ether, and20.0 mL of a 1:1(v/v) mixture of 10% aqueous sodium thiosulfate solutionand saturated aqueous sodium bicarbonate solution was added. Theresultant mixture was extracted with 100.0 mL dichloromethane. Theorganic layer was washed with 50.0 mL saturated aqueous sodium chloridesolution, dried over 50.0 g sodium sulfate, and the solvents evaporatedunder reduced pressure at room temperature. The residue was purified onsix 0.5 mm silica gel preparative thin layer chromatography plates (E.M.Science, Gibbstown, N.J.) using, as the elution solvent, 95:5(v/v)dichloromethane/methyl alcohol. The product bands were eluted from thesilica gel with ethyl acetate to provide 110.0 mg (0.058 mmol) ofCompound 29 {R_(f) : 0.59 [dichloromethane:methyl alcohol, 95:5(v/v)]}in 38.2% yield and 120.0 mg (0.064 mmol) of Compound 30 {R_(f) : 0.53[dichloromethane:methyl alcohol, 95:5(v/v)]} in 42% yield.

To produce Lipid A Analogs B218-31 and B218-32 Compound 29 wasdeprotected generally as described below for the preparation of Compound31. Lipid A Analog B218-32 was produced by reacting the free acidproduct with L-lysine as described below for analog B214-32. Lipid AAnalog B218-31 was produced by reacting the free acid product with Trisas described below for B214-31. ##STR131##

Compound 30 (332.0 mg, 0.176 mmol) was dissolved in 40.0 mL anhydroustetrahydrofuran:96% formic acid (Aldrich Chemical Co.) [10:1(v/v)],under a nitrogen atmosphere in the dark. To this solution was addedtetrakis(triphenylphosphine)palladium(O) (2.07 g, 1.76 mmol (AldrichChemical Co.) and triphenylphosphine (1.45 g, 5.28 mmol, AldrichChemical Co.). The mixture was stirred at room temperature for twohours, the solvents evaporated under reduced pressure at roomtemperature, and the resulting residue azeotroped with 5.0 mL toluenethree times. The residue was then dissolved in 10.0 mL methyl alcohol,and hydrogen sulfide gas was bubbled through the solution for fiveminutes. The solvent was then removed by evaporation under reducedpressure at room temperature. The crude product was purified on aDEAE-cellulose (100.0 g; Sigma Chemical Co., St. Louis, Mo.) columnusing a 0 to 0.1M ammonium acetate (Aldrich Chemical Co.) salt gradientin a 3:2:1(v/v/v) mixture of methyl alcohol/chloroform. Fractionscontaining product (as determined by thin layer chromatographicanalysis) were combined and an equal volume of chloroform added. Theorganic layer was separated and concentrated under reduced pressure atroom temperature, yielding the product in its ammonium salt form. Theproduct was then dissolved in 100.0 mL water, and the excess ammoniumacetate was removed by lyophilization. This product obtained is Lipid AAnalog B214-33.

The lyophilized product was converted to the free acid by passagethrough a CM-cellulose (Sigma Chemical Co., St. Louis, Mo.) column,eluting with 3:2:1(v/v/v) methyl alcohol/chloroform/water. The solutionof free acid product was evaporated to dryness under reduced pressure atroom temperature and an accurate weight obtained.

The product was then dissolved in 5.0 mL methyl alcohol, and 73 mg (0.49mmol) L-lysine (Sigma Chemical Co., "Cell culture grade") dissolved in5.0 mL water was added. The resulting mixture was evaporated to drynessunder reduced pressure at room temperature, the product obtainedredissolved in 300.0 mL pyrogen free deionized water, filtered through a0.2 μm pore size Teflon HPLC filter (Rainin Instruments, Woburn, Mass.),and lyophilized to provide 256.7 mg (0.124 mmol) of a tetralysine salt{i.e., Compound 31; R_(f) : 0.64 [chloroform:methyl alcohol:glacialacetic acid:water, 125:75:10:20(v/v/v/v]} as a white hydroscopic foam in71% yield. This product, Compound 31, is Lipid A Analog, B214-32.

Lipid A Analog B214-31 was produced by reacting the free acid productobtained above with tris[hydroxymethyl]aminomethane (Sigma ChemicalCo.). The resulting mixture was evaporated to dryness under reducedpressure at room temperature, the product obtained redissolved inpyrogen free deionized water, filtered through a 0.2 μm pore size TeflonHPLC filter (Rainin Instruments, Woburn, Mass.), and lyophilized toprovide the tris[hydroxymethyl]amino methane salt, B214-31. ##STR132##

Compound 25 (3.90 g, 2.74 mmol) was dissolved in 40.0 mL anhydrousdichloromethane at room temperature. To this solution, at roomtemperature, was added 3.6 g (10.9 mmol) of Compound C8 (see below) and4.50 g (21.9 mmol) of 1,3-dicyclohexylcarbodiimide. The reaction wasallowed to proceed for 14 hours, at which time the reaction wasdetermined to be complete by thin layer chromtographic analysis[hexanes:ethyl acetate, 1:1(v/v)]. The reaction mixture was then dilutedwith 100.0 mL hexanes and filtered through 20.0 g Celite 545, the solidswashed with 100.0 mL ethyl acetate, and the filtrate evaporated underreduced pressure at room temperature to yield a syrupy residue. Thesyrup was dissolved in 5.0 mL dichloromethane, applied to a silica gel(400.0 g) column and eluted first with 1:4(v/v) ethyl acetate/hexanes,and then with 1:1(v/v) ethyl acetate/hexanes to provide 3.36 g (1.64mmol) of Compound 32 {R_(f) : 0.51 [ethyl acetate:hexanes, 1:1(v/v)]} in60% yield. ##STR133##

Compound 32 (3.46 g, 1.69 mmol) was dissolved in 35.0 mL dichloromethaneand 3.5 mL tert-butyl alcohol. To this solution was added 3.5 mL of pH7.0 phosphate buffer concentrate, followed by 957.0 mg (4.2 mmol) of2,3-dichloro-5,6-dicyano-1, 4-benzoquinone. The resultant heterogeneousmixture was magnetically stirred under a nitrogen atmosphere in the darkfor about 12 hours or until completion [as indicated by thin layerchromatographic analysis using dichloromethane:methyl alcohol,19:1(v/v)]. The reaction was quenched with 30.0 mL 10% aqueous sodiumthiosulfate solution, diluted with 200.0 mL dichloromethane, and pouredinto 100.0 mL saturated aqueous sodium bicarbonate solution. The organiclayer was separated, washed with 100.0 mL saturated aqueous sodiumchloride solution, dried over 100.0 g sodium sulfate, and filteredthrough a cotton plug. The crude reaction mixture was then applieddirectly to a silica gel (400.0 g) column and eluted with adichloromethane:methyl alcohol step gradient [99:1-50:1-19:1-4:1(v/v)].Evaporation of solvent form the product-containing fractions (asdetermined by thin layer chromatographic analysis) under reducedpressure at room temperature provided 2.8 g (1.47 mmol) of Compound 33{R_(f) : 0.32 [dichloromethane:methyl alcohol, 19:1(v/v)]} in 87% yield.##STR134##

To a solution of Compound 33 (716.4 mg, 0.377 mmol) in anhydroustetrahydrofuran (71 mL), 1.1M n-butyllithium (377.0 μL, 0.415 mmol) inhexanes was added slowly with stirring, under a nitrogen atmosphere, at-78° C. After five minutes, 0.5M diallyl chlorophosphate (1.13 mL; 0.566mmol) in anhydrous toluene was added, and the mixture was stirred for 10min. The mixture was then warmed to 0° C., stirred for an additional 10minutes, and quenched with glacial acetic acid (716.0 L). The reactionmixture was poured into 100.0 mL of a saturated aqueous sodiumbicarbonate solution and extracted with 500.0 mL dichloromethane. Theorganic layer was washed first with 100.0 mL saturated aqueous sodiumbicarbonate solution and then with 100.0 mL saturated aqueous sodiumbicarbonate solution and then with 100.0 mL saturated aqueous sodiumchloride solution, dried over 300.0 g sodium sulfate, filtered through acotton plug, and the solvents evaporated under reduced pressure at roomtemperature. The residue obtained was purified by elution from a silicagel (100.0 g) column with 3:1(v/v) toluene/ethyl acetate. Evaporation ofsolvent from the product-containing fractions (as determined by thinlayer chromatographic analysis) under reduced pressure at roomtemperature provided 450.4 mg (0.219 mmol) of Compound 34 {R_(f) : 0.50[dichloromethane:methyl alcohol, 19:1(v/v)]} in 58% yield. ##STR135##

Compound 34 (810.0 mg, 0.40 mmol) was dissolved in acetonitrile (10.0mL), and 1.0 mL water was added. To the resultant solution was addedmercury(II)oxide red (693.0 mg, 3.20 mmol, Aldrich Chemical Co.),followed by mercury(II)chloride (434.4 mg, 1.60 mmol, Aldrich ChemicalCo.), and the resulting mixture stirred at room temperature under anitrogen atmosphere for one hour. The reaction mixture was then dilutedwith 20.0 mL methyl alcohol; hydrogen sulfide was bubbled through themixture for five minutes; and the solution was filtered through a pad ofsilica gel (10.0 g) which had been preconditioned with 4:1(v/v)dichloromethane/methyl alcohol. The filtrate was evaporated to drynessunder reduced pressure at room temperature and purified on a silica gel(100.0 g) column, eluting first with 7:4(v/v) hexanes/diethyl ether andthen with 4:1(v/v) dichloromethane/methyl alcohol. Evaporation ofsolvent from the product-containing fractions (as determined by thinlayer chromatographic analysis) under reduced pressure at roomtemperature provided 539.0 mg (0.287 mmol) of Compound 30 {R_(f) : 0.53[dichloromethane:methyl alcohol, 95:5(v/v)]} in a 72% yield. ##STR136##

To a magnetically stirred solution of Compound 9 (380.0 g; 1.3 mol)dissolved in 1.5 L of N,N-dimethylformamide was first added 227.0 g(3.25 mol) of imidazole under nitrogen, at 0° C., and then added 263.0 g(1.7 mol) of tert-butyldimethylsilyl chloride. The solution was stirredfor one and a half hours, diluted with 2.0 L ethyl acetate and pouredinto 2.0 L saturated aqueous sodium bicarbonate solution. The organiclayer was separated and washed first with 2.0 L saturated aqueous sodiumbicarbonate solution, then with 2.0 L water, and finally with 1.0 Lsaturated aqueous sodium chloride solution. The organic layer was thendried over 500.0 g sodium sulfate, filtered through a glass frittedfunnel and concentrated under reduced pressure, at room temperature. Theresidue was then purified on a silica gel [4.0 kg] column and eluted andethyl acetate:hexanes [1:4(v/v)]. Evaporation of solvent from theproduct-containing fractions (identified by use of thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 411.0 g (1.0mol) of Compound 36 [ethyl acetate:hexanes, 1:4(v/v)]} in 78% yield.##STR137##

To a solution of Compound 36 (411.0 g; 1.0 mol) dissolved in 8.0 Lmethyl alcohol was added 50.0 mL of a 25% (wt/v) sodium methoxide inmethyl alcohol solution, and the resulting mixture was stirred at roomtemperature for six hours. The reaction mixture was then neutralizedwith 1.0 L saturated aqueous ammonium chloride solution and extractedwith 8.0 L ethyl acetate. The organic layer was separated, washed firstwith 1.0 L water, then with 1.0 L saturated aqueous sodium chloridesolution, dried over 1.5 kg sodium sulfate, filtered, and concentratedunder reduced pressure at room temperature. The crude product waspurified on a silica gel [4.0 kg] column and eluted with a stepgradient, beginning with a 5:1(v/v), followed by a 4:1, a 3:1, andfinally with a 2:1 mixture of hexanes:ethyl acetate. Evaporation ofsolvent from the product-containing fractions (as identified by use ofthin layer chromatographic analysis) under reduced pressure at roomtemperature and drying overnight under vacuum at room temperatureprovided 339.8 g (0.95 mol) of Compound 37 [hexanes:ethyl acetate,2:1(v/v)] in a 93% yield. ##STR138##

To a solution of Compound 37 (0.5 g; 1.4 mmol) in 10.0 mL of anhydrousdichloromethane was added: 0.38 g (1.4 mmol) of Compound A6 (see below),0.35 g (1.7 mmol) 1,3-dicyclohexylcarbodiimide, and 1.8 mg (0.02 mmol)of 4-dimethylaminopyridine sequentially, at 0° C., with magneticstirring. The mixture was stirred for an additional three hours, dilutedwith 20.0 mL hexanes, and filtered through 5.0 g Celite 545. Thefiltrate was concentrated under reduced pressure, at room temperature,and the residue purified on a silica gel (100.0 g) column and elutedwith ethyl acetate:hexanes [1:7(v/v)]. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum, at room temperature provided 0.63 g (1.02mmol) of Compound 38 {R_(f) : 0.64 [ethyl acetate:hexanes, 1:4(v/v)]} ina 73% yield. ##STR139##

Compound 38 (0.63 g; 1.02 mmol) was dissolved in 8.0 mL of glacialacetic acid and 2.0 mL of water by magnetic stirring at room temperaturefor 12 hours. The mixture was concentrated under reduced pressure atroom temperature and azeotroped three times with 10.0 mL portions oftoluene. The residue was purified on a silica gel (100.0 g) column andeluted with 1:2(v/v) ethyl acetate:hexanes. Evaporation of solvent fromthe product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 0.57 g (0.99mmol) of Compound 39 {R_(f) : 0.22 [ethyl acetate:hexanes, 1:2(v/v)]} in98% yield. ##STR140##

A mixture of Compound 23A (113.4 mg; 0.134 mmol) and Compound 39 (321.5mg; 0.61 mmol) was dried under vacuum for 14 hour and dissolved in 10.0mL of anhydrous toluene. To this solution was added 300.0 mg of powderedAW-300 molecular sieves which were flame-dried under vacuum, and theresulting mixture magnetically stirred for one hour at room temperatureunder an argon atmosphere. The mixture was then cooled to -35° C. and8.0 mL (0.32 mmol) of a 0.04M boron trifluoride etherate:toluenesolution [prepared by dissolving 200.0 μL (1.6 mmol) of borontrifluoride etherate in 40.0 mL of toluene and stirring with 200.0 mgpowdered AW-300 molecular sieves for one hour at room temperature] wasadded over a two and a half hour period using a syringe pump. Thereaction was quenched with 10.0 mL of saturated aqueous sodiumbicarbonate solution, diluted with 100.0 mL dichloromethane, andfiltered through 20.0 g Celite 545. The filtrate was washed first with100.0 mL of water, and finally with 100 mL of saturated aqueous sodiumchloride solution, dried over 50.0 g sodium sulfate, filtered though aglass fritted funnel, and concentrated under reduced pressure at roomtemperature. The residue was purified on a silica gel (100.0 g) columnand eluted with ethyl acetate:hexanes [1:2(v/v)]. Evaporation of solventfrom the product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 119.1 g(0.094 mmol) of Compound 40 {R_(f) : 0.44 [ethyl acetate:hexanes,1:2(v/v)]} in 70% yield. ##STR141##

To a magnetically stirred solution of Compound 40 (110.0 mg; 0.09 mmol)dissolved in 1.0 mL anhydrous toluene and 33.0 μL (0.34 mmol) anhydrouspyridine, at 0° C., was added 68.0 μL (0.13 mmol) 1.93M phosgene intoluene dropwise, and the reaction mixture was stirred for an additional15 minutes. To this solution, 100.0 μL (1.47 mmol) of allyl alcohol wasadded dropwise, the reaction was stirred for an additional 30 minutes,the reaction quenched by addition, at 0° C., of 10.0 mL saturated sodiumbicarbonate solution and warming to 25° C. The reaction mixture was thenextracted with 100.0 mL ethyl acetate, the organic layer washed with10.0 mL saturated aqueous sodium chloride solution, dried over 25.0 gsodium sulfate, filtered through a glass fritted funnel, andconcentrated under reduced pressure at room temperature. The residueobtained was purified on a silica gel (100.0 g) column and eluted with1:3(v/v) mixture of ethyl acetate:hexanes. Evaporation of solvent fromthe product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 75.0 mg (0.06mmol) of Compound 41 {R_(f) : 0.75 [hexanes:ethyl acetate, 3:1(v/v)]} in64% yield. ##STR142##

To a magnetically stirred solution of Compound 41 (75.0 mg; 0.056 mmol)dissolved in 4.0 mL of anhydrous dichloromethane, was added 250.0 mg(0.45 mmol) of tin(II)tris-benzenethiolate triethylamine complex and theresulting mixture stirred at room temperature under nitrogen in the darkuntil thin layer chromatographic analysis [hexanes:ethyl acetate,1:1(v/v)] indicated the starting material to be consumed (i.e., for twohours). The reaction mixture was loaded directly onto a silica gel (10.0g) column and eluted first with a 4:1(v/v) mixture of hexanes:ethylacetate and then with ethyl acetate. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying under vacuum at room temperature for 30 minutes provided 64.4 mg(0.05 mmol) of Compound 42 {R_(f) : 0.57 [ethyl acetate:hexanes,1:1(v/v)]} in 90% yield. ##STR143##

To a magnetically stirred 0° C. solution of Compound 42 (64.4 mg; 0.05mmol) in 3.0 mL anhydrous dichloromethane was added 67.0 mg (0.28 mmol)of compound D2 and 70.0 mg (0.34 mmol) of 1,3-dicyclohexylcarbodiimide.After one hour, when thin layer chromatographic analysis [hexanes:ethylacetate, 1:1(v/v)] indicated completion of the reaction had occurred,the reaction mixture was diluted with 50.0 mL ethyl acetate and filteredthrough 10.0 g Celite 545. The solids obtained were washed with 20.0 mLethyl acetate and the filtrate concentrated under reduced pressure atroom temperature yielding a syrupy residue. The syrup was dissolved in1.0 mL dichloromethane, loaded onto a silica gel (10.0 g) column andeluted first with 1:9(v/v) ethyl acetate:hexanes to remove reagentresidues, and then with 1:1(v/v) ethyl acetate:hexanes. Evaporation ofsolvent from the product-containing fractions (as indicated by thinlayer chromatographic analysis) under reduced pressure at roomtemperature and drying overnight under vacuum at room temperatureprovided 87.0 mg (0.04 mmol) of Compound 43 {R_(f) : 0.95 [ethylacetate:hexanes, 1:1(v/v)]} in 85% yield. ##STR144##

To a magnetically stirred solution of 12.0 mL of 2M hydrogen fluoride inacetonitrile in a Teflon reaction vessel was added 70.0 mg (0.036 mmol)of Compound 43 dissolved in 0.5 mL of dichloromethane, at roomtemperature. The resulting mixture was stirred for 18 additional hours,diluted with 20.0 mL saturated aqueous sodium bicarbonate solution, andextracted with 100.0 mL dichloromethane. The organic layer was separatedand washed first with 20.0 mL water and then with 10.0 mL saturatedaqueous sodium chloride solution. The organic layer was dried over 25.0g sodium sulfate, filtered through a glass fritted funnel, andconcentrated under reduced pressure at room temperature. The residue waspurified on a silica gel (10.0 g) column and eluted withdichloromethane:methyl alcohol [98:2(v/v)]. Evaporation of solvent fromthe product-containing fractions (as indicated by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 60.3 mg(0.035 mmol) of Compound 44 {R_(f) : 0.78 [dichloromethane:methylalcohol, 98:2(v/v)]} in 97% yield.

To produce Lipid A Analog B276-32, Compound 44 was deprotected generallyas described above for the preparation of Compound 31, and the free acidproduct was reacted with L-lysine as described above for analog B214-32.##STR145##

To a magnetically stirred solution of Compound 44 (65.0 mg; 0.038 mmol)in 5.0 mL anhydrous dichloromethane, 70.0 mg (0.28 mmol) ofbis(allyloxy)(diisopropylamino) phosphine and 70.0 mg (1.0 mmol) of1H-tetrazole was added at 0° C., under a nitrogen atmosphere. Themixture was warmed to room temperature and stirred for an additionalhour. The mixture was then cooled to -78° C., and a solution of 11.95 mg(0.12 mmol) 3-chloroperoxybenzoic acid dissolved in 0.80 mLdichloromethane was added. The mixture was warmed to 0° C., stirred form20 additional minutes, and 10.0 mL saturated aqueous sodium bicarbonatesolution was added. The resultant mixture extracted with 100.0 mLdichloromethane, and the organic layer was separated and washed firstwith 10.0 mL water, and then with 10.0 mL saturated aqueous sodiumchloride solution, and dried over 25.0 g sodium sulfate. The driedproduct was concentrated under reduced pressure, at room temperaturepurified on a silica gel (10.0 g) column, and eluted with ethylacetate:hexanes [1:1(v/v)]. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 53.0 mg(0.028 mmol) of Compound 30 {R_(f) : 0.29 [ethyl acetate:hexanes,1:1(v/v)]} in a 74% yield. ##STR146##

To a magnetically stirred solution of Compound 37 (19.0 g; 0.05 mol)dissolved in 65.0 mL anhydrous dichloromethane, under an argonatmosphere, at 0° C. was added 2.75 g (0.11 mol) 60% sodium hydride oildispersion (Aldrich Chemical. Co.). The mixture was stirred first forfive minutes at 0° C. and then for 15 minutes at room temperature. Underargon, a solution of 20.5 g (0.06 mol) A10 dissolved in 30.0 mL ofanhydrous dichloromethane was then added dropwise to the reactionmixture through a syringe-pump over a two-hour period. After stirringfor 30 minutes, the reaction mixture was cooled to 0° C., 5.0 mL methylalcohol was added dropwise to quench any unreacted sodium hydride, andthe reaction mixture was diluted with 300.0 mL dichloromethane andwashed first with 300.0 mL saturated aqueous ammonium chloride solution,then with 300.0 mL saturated aqueous sodium chloride solution. Theorganic layer was separated, dried over 100.0 g sodium sulfate andconcentrated under reduced pressure to yield a crude syrupy product. Theproduct was purified on silica gel (2.0 kg) column and eluted with astep gradient of hexanes:ethyl acetate [12:1 to 9:1 to 8:1 to 5:1(v/v)].Evaporation of solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 20.1 g (0.04 mol) of Compound 45 [R_(f) : 0.53[hexanes:ethyl acetate, 4:1(v/v)]} in 68% yield. ##STR147##

To a magnetically stirred solution of Compound 45 (13.69 g; 26.5 mmol)and Compound B4 (6.3 g; 31.8 mmol) see below) dissolved in 76.0 mL ofanhydrous dichloromethane at room temperature under a nitrogenatmosphere was added 11.0 g (53.0 mmol) of 1,3-dicyclohexylcarbodiimde.To the resulting mixture was added 5.26 mL (0.26 mmol) of 0.5M4-dimethylaminopyridine in anhydrous dichloromethane over a one-hourperiod. The reaction mixture was stirred for nine hours at roomtemperature, filtered through a pad of 100.0 g Celite 545, and thefiltered solids washed with 200.0 mL of ethyl acetate. The combinedfiltrate and ethyl acetate wash were concentrated under reduced pressureat room temperature and the crude product purified on a silica gel (2.0kg) column and eluted with hexanes:ethyl acetate [9:1(v/v)]. Evaporationof solvent from the product-containing fractions (as identified by thinlayer chromatographic analysis) under reduced pressure at roomtemperature and drying overnight under vacuum at room temperatureprovided 17.4 g (25.1 mmol) of Compound 46 {R_(f) : 0.80 [hexanes:ethylacetate, 4:1(v/v)]} in 94% yield. ##STR148##

A solution of Compound 46 (17.4 g; 25.0 mmol), dissolved in 100.0 mL ofa 8:1(v/v) mixture of glacial acetic acid and water, was heated to 60°C., with magnetic stirring for 12 hours. The reaction mixture was thenconcentrated under reduced pressure, at 40° C., and the crude productpurified on a silica gel (2.0 kg) column by elution with a step gradientof hexanes:ethyl acetate [first 6:1(v/v) then 2:1(v/v)]. Evaporation ofsolvent from the product-containing fractions (as identified by thinlayer chromatographic analysis) under reduced pressure at roomtemperature and drying overnight under vacuum at room temperatureprovided 15.0 g (22.9 mmol) of Compound 47 {R_(f) : 0.13 [hexanes:ethylacetate, 4:1(v/v)]} in 91% yield. ##STR149##

To a magnetically stirred solution of Compound 47 (8.85 g; 12.7 mmol)dissolved in 60.0 mL of a 4:1[v/v] mixture of anhydrous toluene andanhydrous pyridine, at 0° C., under nitrogen, was added 1.75 mL (16.5mmol) allychloroformate dropwise over a 30-minute period. The resultingmixture was diluted with 300.0 mL ethyl acetate, washed with 100.0 mLsaturated aqueous sodium bicarbonate solution, 100 mL water, and 100 mLsaturated aqueous sodium chloride solution, dried over 100.0 g sodiumsulfate, and concentrated under reduced pressure at room temperature.The residue was dissolved in 10.0 mL dichloromethane, loaded on a silicagel (1.0 kg) column and eluted with ethyl acetate:hexanes [1:9(v/v)].Evaporation of solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 8.1 g (10.9 mmol) of Compound 48. ##STR150##

To a magnetically stirred solution of Compound 48 (1.6 g; 2.06 mmol),dissolved in 10.0 mL anhydrous dichloromethane at room temperature undera nitrogen atmosphere, was first added 757.0 mg (3.1 mmol)bis(allyloxy)(diisopropylamino) phosphine and then added (in oneportion) 650.0 mg (9.3 mmol) 1H-tetrazole. After 10 minutes, thereaction mixture was cooled to -78° C., and a solution of 550.0 mg (2.2mmol) of 55% 3-chloroperoxybenzoic acid dissolved in 5.0 mL anhydrousdichloromethane was added dropwise over a 10-minute period. The reactionwas quenched at -78° C. by the addition of 50.0 mL saturated aqueoussodium bicarbonate solution. The resulting mixture was then extractedwith 200.0 mL dichloromethane and the organic layer extracted washedfirst with 50.0 mL water, then with 50.0 mL saturated aqueous sodiumchloride, and dried over 50.0 g sodium sulfate. Concentration underreduced pressure at room temperature provided the crude product whichwas purified on a silica gel (300.0 g) column and eluted with ethylacetate:hexanes [1:4(v/v)]. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 1.7 g (1.8mmol) of Compound 49. ##STR151##

To a magnetically stirred solution of 70.0 mL of 6M hydrogen fluoride inacetonitrile in a Teflon reaction vessel was added 10.5 g (11.6 mol) ofCompound 49 dissolved in 10.0 mL of dichloromethane at room temperature.The resulting mixture was stirred for an additional 18 hours, pouredinto 400.0 mL of a saturated aqueous sodium bicarbonate solution at 0°C., and extracted with 500.0 mL dichloromethane. The organic layerextract was washed first with 100.0 mL water and then with 100.0 mLsaturated aqueous sodium chloride solution. The organic layer was driedover 250.0 g sodium sulfate, filtered through a glass fritted funnel,and concentrated under reduced pressure at room temperature. The residuewas purified on a silica gel (1.0 kg) column and eluted withhexanes:ethyl acetate [3:1(v/v)]. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 7.9 g (10.1mmol) of Compound 50. ##STR152##

To a mechanically stirred solution of Compound 50 (1.1 g; 1.4 mmol) in10.0 mL trichloroacetonitrile, 2.6 g (8.0 mmol) of cesium carbonate wasadded at room temperature under a nitrogen atmosphere. After two hours,the mixture was filtered through 25.0 g Celite 545, the filtered solidswashed with 100.0 mL dichloromethane, and the combined filtratesconcentrated under reduced pressure at room temperature. The crudeproduct was purified on a silica gel (200.0 g) column and eluted withdichloromethane:diethyl ether [19:1(v/v)]. Evaporation of solvent fromthe product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 600.7 mg(0.65 mmol) of Compound 51A (α isomer) and 500.0 mg (0.54 mmol) ofCompound 51B (β isomer) in a combined yield of 85%. ##STR153##

To a magnetically stirred solution of Compound 45 (13.8 g; 26.8 mmol)dissolved in 100.0 mL of a 4:1(v/v) mixture of anhydrous toluene andanhydrous pyridine at room temperature under a nitrogen atmosphere, wasadded 21.0 mL (40.2 mmol) of 1.93M phosgene in toluene dropwise over a30-minute period. The resulting mixture was stirred for an additional 15minutes, 16.1 mL (214.4 mmol) allyl alcohol was added and the mixture(214.4 mmol) allyl alcohol was reaction mixture was diluted with 100.0mL saturated aqueous sodium bicarbonate solution and extracted with300.0 mL ethyl acetate. The organic layer extract was washed first with200.0 mL water, then with 100.0 mL saturated aqueous sodium chloridesolution, and dried over 200.0 g sodium sulfate. The dried organic layerextract was filtered and concentrated under reduced pressure, at roomtemperature. The crude product obtained was dissolved in 10.0 mLdichloromethane, loaded onto a silica gel (1.0 kg) column and elutedwith ethyl acetate:hexanes [1:9(v/v)]. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 15.6 g (26.1mmol) of Compound 52 in 97% yield. ##STR154##

A solution of Compound 52 (15.6 g; 26.1 mmol) dissolved in 50.0 mL ofglacial acetic acid and 2.0 mL of water was magnetically stirred at roomtemperature for 12 hours. The mixture was concentrated under reducedpressure at room temperature and azeotroped three times with 10.0 mLportions of toluene. The residue was purified on a silica gel (1.0 kg)column and eluted using a two step gradient: 1:2(v/v) ethylacetate:hexanes then ethyl acetate. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 12.1 g (21.6mmol) of Compound 53 in 83% yield. ##STR155##

To a magnetically stirred solution of Compound 53 (10.3 g; 18.5 mmol)dissolved in 400.0 mL of anhydrous dichloromethane, under a nitrogenatmosphere, at 0° C., was added 2.9 g (42.6 mmol) of imidazole, followedby 3.6 g (24.1 mmol) of tert-butyldimethylsilyl chloride. The resultingmixture was warmed to room temperature and stirred for three hours. Thereaction mixture was poured into 1.0 L saturated aqueous ammoniumchloride solution, and the product extracted with 1.0 L dichloromethane.The organic layer extract was washed first with 200.0 mL saturatedaqueous sodium bicarbonate solution, then with 200.0 mL water, andfinally with 100.0 mL saturated aqueous sodium chloride solution. Thewashed organic layer was dried over 300.0 g sodium sulfate, filtered andconcentrated under reduced pressure, at room temperature. The crudeproduct was purified on a silica gel (1.0 kg) column and eluted withethyl acetate:hexanes [1:8(v/v)]. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 10.6 g (15.8mmol) of Compound 54 {R_(f) : 0.70 [ethyl acetate:hexanes, 1:4(v/v)]} in85% yield. ##STR156##

To a mechanically stirred solution of Compound 54 (8.9 g; 13.2 mmol)dissolved in 270.0 mL of anhydrous toluene and 4.2 mL of anhydrouspyridine, at 0° C., under a nitrogen atmosphere, was slowly added 10.2mL (26.4 mmol) of 1.93M phosgene in toluene, over a 10-minute period.Twenty minutes later, 8.0 mL (105.6 mmol) of allyl alcohol was addedover a five-minute period, and the resulting reaction mixture stirredfor an additional 15 minutes. The reaction mixture was quenched with200.0 mL saturated aqueous sodium bicarbonate solution, diluted with 1.0L ethyl acetate, and the organic layer separated, and washed with 500.0mL water and then 500.0 mL saturated aqueous sodium chloride solution,dried over 500.0 g sodium sulfate, filtered, and concentrated underreduced pressure at room temperature. The residue was purified on asilica gel (1.0 kg) column and eluted with ethyl acetate:hexanes[1:19(v/v)]. Evaporation of solvent from the product-containingfractions (as identified by thin layer chromatographic analysis) underreduced pressure at room temperature and drying overnight under vacuumat room temperature and drying overnight of Compound 55 {R_(f) : 0.68[ethyl acetate:hexanes, 1:9(v/v)]} in 95% yield. ##STR157##

In a 1.0 L Teflon reaction vessel, Compound 55 (5.8 g; 7.6 mmol) wasdissolved in 200.0 mL of dichloromethane. To the solution at roomtemperature, with magnetic stirring, was added 150.0 mL of a 1M solutionof hydrofluoric acid in acetonitrile. After seven hours, the reactionmixture was quenched by pouring into 200.0 mL saturated aqueous sodiumbicarbonate solution, at 0° C., and extracted with 500.0 mLdichloromethane. The organic layer was separated, washed with 100.0 mLwater then with 100.0 mL saturated aqueous sodium chloride solution,dried over 300.0 g sodium sulfate, filtered, and concentrated underreduced pressure at room temperature. The residue obtained was purifiedon a silica gel [600.0 g] column and eluted with ethyl acetate:hexanes[1:4(v/v)]. Evaporation of solvent from the product-containing fractions(as identified by a thin layer chromatographic analysis) under reducedpressure at room temperature provided 4.5 g (6.7 mmol) of Compound 56{R_(f) : 0.33 [ethyl acetate:hexanes, 1:4(v/v)]} in 88% yield.##STR158##

A heterogeneous mixture of 8.0 g (12.2 mmol) Compound 47, 11.3 g (48.8mmol) silver(I) oxide (Aldrich Chemical Co.), and 120.0 mL (1.92 mol)methyl iodide (Aldrich Chemical Co.) was mechanically stirred at 39° C.,for 12 hours under nitrogen in the dark. The reaction mixture wascooled, filtered through a 100.0 g Celite 545 and the filtered solidswashed with 200.0 mL ethyl acetate. The combined filtrate and wash wasthen concentrated under reduced pressure at 40° C., yielding crudeproduct which was then dissolved in 50.0 mL dichloromethane and cooledto 0° C. To the cooled reaction mixture was added, in one portion, 1.0 g(14.69 mmol) imidazole followed by tert-butylchlorodiphenylsilane over afive-minute period. The reaction mixture was then warmed to roomtemperature, stirred one hour longer, quenched with 100.0 mL saturatedaqueous sodium bicarbonate solution, and extracted with 500.0 mL ofdichloromethane. The organic layer was washed first with 100.0 mL waterand then with 100.0 mL saturated aqueous sodium chloride solution, driedover 300.0 g sodium sulfate, filtered and concentrated under reducedpressure at room temperature. The residue obtained was purified on asilica gel (100.0 g) column by elution with ethyl acetate:hexanes[1:9(v/v)]. Evaporation of solvent from the product-containing fractions(identified by use of thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature yielded 6.85 g (10.2 mmol) of Compound 57A {R_(f) : 0.63[dichloromethane:diethyl ether, 19:1(v/v)]} in 84% yield and 1.11 g(1.22 mmol) of Compound 57B {R_(f) : 0.90 [dichloromethane:diethylether, 19:1(v/v)]} in 10% yield. ##STR159##

To a magnetically stirred solution of Compound 57A (8.7; 0.013 mol)dissolved in 46.0 mL anhydrous dichloromethane, at room temperature,under a nitrogen atmosphere, was first added 4.8 mL (0.02 mol)bis(allyloxy)(diisopropylamino)phosphine and then added (in one portion)4.1 g (0.06 mol) 1H-tetrazole. After five minutes, the reaction mixturewas cooled to -78° C., and a solution of 3.35 g (0.02 mol) 55%3-chloroperoxybenzoic acid dissolved in 37.0 mL anhydrousdichloromethane was added dropwise over a 10 minute period. The reactionwas then quenched at -78° C. by the addition of 100.0 mL saturatedaqueous sodium bicarbonate solution. The resulting mixture was thenextracted with 500.0 mL dichloromethane and the organic layer extractwashed first with 200.0 mL water, then with 200.0 mL saturated aqueoussodium chloride solution, and dried over 300.0 g sodium sulfate.Concentration under reduced pressure at room temperature provided thecrude product which was purified on a silica gel (1.0 kg) column andeluted with ethyl acetate:hexanes [1:6(v/v)]. Evaporation of solventfrom the product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 8.8 g (0.11mol) of Compound 58 {R_(f) : 0.28 [ethyl acetate:hexanes, 1:4(v/v)]} inan 85% yield. ##STR160##

To a magnetically stirred solution of 80.0 mL 6M hydrogen fluoride inacetonitrile in a Teflon reaction vessel was added at room temperature,8.8 g (10.6 mmol) of Compound 58 dissolved in 30.0 mL ofdichloromethane. The resulting mixture was stirred for nine hours,poured into 200.0 mL saturated aqueous sodium bicarbonate solutionaqueous sodium chloride solution, dried over 100.0 g sodium sulfate,filtered and concentrated under reduced pressure at room temperature.The residue was then purified on a silica gel (1.0 kg) column and elutedwith hexanes:ethyl acetate [1:1(v/v)]. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 5.7 g (7.95mmol) of Compound 59 {R_(f) : 0.37 [dichloromethane:methyl alcohol,95:5(v/v)]} in 75% yield. ##STR161##

To a mechanically stirred solution of 10.32 g (14.5 mmol) of Compound 59in 200.0 mL trichloroacetonitrile, 8.80 g (63.7 mmol) of potassiumcarbonate was added at room temperature under nitrogen. After 20minutes, the mixture was filtered through 100.0 g Celite 545, thefiltered solids washed with 100.0 mL dichloromethane and the combinedfiltrates concentrated under reduced pressure at room temperature. Thecrude product obtained was purified on a silica gel (10.0 g) column byelution with hexanes:ethyl acetate [1:1(v/v)]. Evaporation of solventfrom the product-containing fractions (identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature gave 11.1 g (12.9mmol) of 60B (β isomer) and 60A (α isomer) {R_(f) : 0.61 and 0.53[hexanes:ethyl acetate, 1:1(v/v)]} in a combined yield of 89%.##STR162##

A mixture of 465.0 mg (0.492 mmol) of Compound 51A and 374.0 mg (0.541mmol) of Compound 17 was dried under vacuum for 14 hours, dissolved in10.0 mL of anhydrous dichloromethane, and to the solution was added800.0 mg of powdered AW-300 molecular sieves, which had been flame-driedunder vacuum. The resulting mixture was magnetically stirred for onehour at room temperature under an argon atmosphere, cooled to -23° C.,and 740.0 μL (0.147 mmol) of a 0.2M boron trifluoride etherate:anhydrousdichloromethane solution [prepared by dissolving 250.0 μL (2.03 mmol) ofboron trifluoride etherate in 10.0 mL of anhydrous dichloromethane andstirring with 200 mg powdered AW-300 molecular sieves for one hour atroom temperature] was slowly added over a one hour period. The reactionwas quenched with 5.0 mL of saturated aqueous sodium bicarbonatesolution, diluted with 100.0 mL dichloromethane, and filtered through10.0 g Celite 545. The filtrate was washed first with 50.0 mL saturatedaqueous sodium bicarbonate solution, then with 50 mL of water, andfinally with 50 mL of saturated aqueous sodium chloride solution, driedover 25.0 g sodium sulfate, filtered, and concentrated under reducedpressure at room temperature. The resulting residue was purified on asilica gel (100.0 g) column and eluted with ethyl acetate:hexanes[1:3(v/v)]. Evaporation of solvent from the product-containing fractions(as identified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 430.0 mg (0.292 mmol) of Compound 62 {R_(f) : 0.2[ethyl acetate:hexanes, 1:2(v/v)]} in 60% yield. ##STR163##

A mixture of 250.0 mg (0.265 mmol) of Compound 23A and 205.0 mg (0.265mmol) of Compound 14 was dried under vacuum for 14 hours and dissolvedin 15.0 mL of anhydrous dichloromethane. To this solution was added600.0 mg of powdered AW-300 molecular sieves, which had been flame-driedunder vacuum, and the resulting mixture magnetically stirred for onehour at room temperature under an argon atmosphere. The mixture wascooled to -23° C. and 400.0 μL (0.265 mmol) of a 0.2M boron trifluorideetherate:anhydrous dichloromethane solution [prepared by dissolving250.0 μL (2.03 mmol) of boron trifluoride etherate in 10.0 mL ofanhydrous dichloromethane and stirring with 200 mg powdered AW-300molecular sieves for one hour at room temperature] was slowly added overa 30-minute period. The reaction was quenched with 5.0 mL of saturatedaqueous sodium bicarbonate solution, diluted with 100.0 mLdichloromethane, and filtered through 10.0 g Celite 545. The filtratewas washed first with 50.0 mL of saturated aqueous sodium bicarbonatesolution, then with 50 mL of water, and finally with 50 mL of saturatedaqueous sodium chloride solution, dried over 25.0 g sodium sulfate,filtered, and concentrated under reduced pressure at room temperature.The resulting residue was purified on a silica gel (200.0 g) column andeluted with ethyl acetate and hexanes [1:3(v/v)]. Evaporation of solventfrom the product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum, at room temperature provided 210.0 mg(0.152 mmol) of Compound 63 {R_(f) : 0.23 [ethyl acetate:hexanes,1:2(v/v)]} in 62% yield. ##STR164##

A mixture of 601.4 mg (0.636 mmol) of Compound 51A and 769.3 mg (1.38mmol) of Compound 39 were dried under vacuum for 14 hours and dissolvedin 40.0 mL of anhydrous toluene. To this solution was added 1.0 g ofpowdered AW-300 molecular sieves, which had been flame-dried undervacuum, and the resulting mixture magnetically stirred for one hour atroom temperature under an argon atmosphere. The mixture was cooled to-35° C. and 10.0 mL (0.190 mmol) of a 0.02M boron trifluorideetherate:anhydrous dichloromethane solution [prepared by dissolving250.0 μL (2.03 mmol) of boron trifluoride etherate in 10.0 mL ofanhydrous dichloromethane, diluting the resulting mixture with 91.5 mLanhydrous toluene and stirring with 200 mg powdered AW-300 molecularsieves for one hour at room temperature] was slowly added over a one anda half hour period. The reaction was quenched with 10.0 mL of saturatedaqueous sodium bicarbonate solution, diluted with 200.0 mLdichloromethane, and filtered through 10.0 g Celite 545. The filtratewas washed first with 50.0 mL of saturated aqueous sodium bicarbonatesolution, then with 50 mL of water, and finally with 50 mL of saturatedaqueous sodium chloride solution, dried over 50.0 g sodium sulfate,filtered, and concentrated under reduced pressure at room temperature.The resulting residue was purified on a silica gel (100.0 g) column andeluted with ethyl acetate:hexanes [1:2(v/v)]. Evaporation of solventfrom the product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 297.5 mg(0.152 mmol) of Compound 64 {R_(f) : 0.42 [dichloromethane:diethylether, 9:1(v/v)]} in 34% yield. ##STR165##

A mixture of 7.35 g (8.5 mmol) of Compound 60 (αβ mixture) and 5.0 g(7.4 mmol) of Compound 56 which had been dried under vacuum for 14hours, was dissolved in 200.0 mL of anhydrous dichloromethane. To thissolution was added 8.2 g of powdered AW-300 molecular sieves (previouslyflame-dried under vacuum) and the resulting mixture magnetically stirredfor one hour at room temperature under argon. The mixture was thencooled to -35° C., and 8.7 mL (0.50 mmol) of a 0.05M trimethylsilyltrifluoromethanesulfonate (Aldrich Chemical Co.):dichloromethanesolution [prepared by dissolving 310.0 μL (2.03 mmol) oftrimethylsilylmethyl trifluoromethanesulfonate in 40.0 mL of anhydrousdichloromethane and stirring with 1.0 g powdered AW-300 molecular sievesfor one hour at room temperature] was slowly added over an eight hourperiod. The reaction was quenched with 100.0 mL of saturated aqueoussodium bicarbonate solution, then diluted with 500.0 mL dichloromethane,and filtered through 50.0 g Celite 545. The filtrate was then washedwith 100.0 mL portions of saturated aqueous sodium bicarbonate solution,water, and saturated aqueous sodium chloride solution sequentially,dried over 100.0 g sodium sulfate, filtered, and then concentrated underreduced pressure at room temperature. The resulting residue was purifiedon a silica gel (200.0 g) column by elution with ethyl acetate andhexanes [1.4(v/v)]. Evaporation of solvent from the product containingfractions (identified by use of thin layer chromatography analysis)under reduced pressure at room temperature and drying overnight undervacuum at room temperature gave 8.1 g (0.006 mol) of Compound 65 {R_(f): 0.42 [ethyl acetate:hexanes, 1:2(v/v)]} in 82% yield. ##STR166##

To a magnetically stirred solution of Compound 65 (1.99 g; 1.48 mmol)dissolved in 10.0 mL of anhydrous dichloromethane was added 250.0 mg(0.45 mmol) of tin(II)tris-benzenethiolate triethylamine complex and theresulting mixture stirred at room temperature under a nitrogenatmosphere in the absence of light of 30 minutes, at which time thinlayer chromatographic analysis [hexanes:ethyl acetate, 1:1(v/v)]indicated starting material to be consumed. The reaction mixture wasloaded directly into a silica gel (10.0 g) column and eluted first witha 4:1(v/v) mixture of hexanes:ethyl acetate to remove reagentby-products and then with ethyl acetate. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying under vacuum at room temperature for 30 minutes providedpartially purified Compound 66 [1.72 g (1.33 mmol)] {R_(f) : 0.48[dichloromethane:methyl alcohol, 95:5(v/v)]} which was suitable for usein subsequent reaction in 90% yield. ##STR167##

To a magnetically stirred solution of Compound 66 (1.72 g; 1.33 mmol) in10.0 mL anhydrous dichloromethane, at 0° C., was added 1.1 g (4.44 mmol)of Compound D2 (see below) and 1.83 g (8.88 mmol) of1,3-dicyclohexylcarbodiimide. After 30 minutes, when thin layerchromatographic analysis [dichloromethane:methyl alcohol, 95:5(v/v)]indicated that completion of the reaction had occurred, the reactionmixture was diluted with 50.0 mL ethyl acetate, filtered through 10.0 gCelite 545, the solids washed with 20.0 mL ethyl acetate, and thefiltrate concentrated under reduced pressure at room temperature toyield a syrupy residue. The crude syrup was dissolved in 5.0 mLdichloromethane, loaded onto a silica gel (100.0 g) column and elutedinitially with a 1:4(v/v) mixture of ethyl acetate:hexanes to removereagent residues and then with a 1:2(v/v) mixture of ethylacetate:hexanes. Evaporation of solvent from the product-containingfractions (as identified by thin layer chromatographic analysis) underreduced pressure at room temperature and drying overnight under vacuumat room temperature provided 1.82 g (1.04 mmol) of Compound 67 {R_(f) :0.54 [dichloromethane:methyl alcohol, 95:5(v/v)]} in 71% yield.##STR168##

To a magnetically stirred solution of 8.0 ml 6M hydrogen fluoride inacetonitrile in a Teflon reaction vessel was added 390.0 mg (0.224 mmol)of Compound 67 dissolved in 0.5 mL of dichloromethane, at roomtemperature. The mixture was stirred for one and a half hours, dilutedwith 20.0 mL saturated aqueous sodium bicarbonate solution, andextracted with 100.0 mL dichloromethane. The organic layer extract waswashed first with 20.0 mL water, and then with 10.0 mL saturated aqueoussodium chloride solution, dried over 25.0 g sodium sulfate, filtered andconcentrated under reduced pressure at room temperature. The residue waspurified on a silica gel [50.0 g] column and eluted withdichloromethane:methyl alcohol [98:2 (v/v)]. Evaporation of solvent fromthe product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 325.0 mg(0.20 mmol) of Compound 68 {R_(f) : 0.52 [dichloromethane and methylalcohol, 95:5 (v/v)]} in 89% yield. ##STR169##

To a magnetically stirred solution of 50.0 mg (0.03 mmol) of Compound 68in 1.0 mL anhydrous dichloromethane was first added 11.3 mg (0.045 mmol)bis(allyloxy) (diisopropylamino) phosphine followed by 9.4 mg (0.135mmol) 1H-tetrazole at 0° C., under a nitrogen atmosphere. The resultingmixture was warmed to room temperature, stirred 20 minutes, cooled to-78° C., and a solution of 9.5 mg (0.036 mmol) 3-chloroperoxybenzoicacid dissolved in 100.0 μL dichloromethane was added, and the mixturewas stirred for 20 additional minutes. A 0.5 mL saturated aqueous sodiumbicarbonate solution was then added, and the resultant mixture extractedwith 10.0 mL dichloromethane. The organic layer was separated and washedfirst with 10.0 mL water and then with 5.0 mL saturated aqueous sodiumchloride solution, and dried over 5.0 g sodium sulfate. Concentration ofthe dried organic extract under reduced pressure at room temperatureprovided the crude product which was purified on a silica gel (10.0 g)column and eluted with ethyl acetate:chloroform [1:1 (v/v)]. Evaporationof solvent from the product-containing fractions (as identified by thinlayer chromatographic analysis) under reduced pressure at roomtemperature and drying for one hour under vacuum at room temperatureprovided 41.7 mg (0.023 mmol) of Compound 69 {R_(f) : 0.40[dichloromethane:methyl alcohol, 95:5 (v/v)]} in a 78% yield. ##STR170##

To a solution of Compound 69 (130.0 mg, 0.072 mmol) dissolved in 10.0 mLtetrahydrofuran:96% formic acid [10:1v/v)], under a nitrogen atmosphere,in the absence of light, was added 843.0 mg (0.72 mmol) tetrakis(triphenylphosphine)palladium(O) and 575.0 mg (2.19 mmol)triphenylphosphine. The resulting mixture was stirred for a total of onehour, and concentrated under reduced pressure at room temperature. Theresulting residue was mixed with 5.0 mL of toluene and evaporated underreduced pressure at room temperature to a thick paste, suspended in 10.0mL methyl alcohol, and hydrogen sulfide gas bubbled through the solutionfor several minutes. The solvent was removed by evaporation underreduced pressure at room temperature and the crude product was taken upin 10.0 mL of a 3:2:1 (v/v/v) mixture of methyl alcohol:chloroform:waterand filtered through a 0.2 μ Teflon HPLC filter (Rainin Instrument Co.).The filtrate was loaded onto a DEAE-celluose [100.0 g (Sigma ChemicalCo.)] column and eluted with 2.0 L of a 3:2:1 (v/v/v) mixture of methylalcohol:chloroform:water, using a 0 to 0.1M ammonium acetate linear saltgradient. The purified product-containing fractions (as identified bythin layer chromatographic analysis) were combined and an equal volumeof chloroform was added. The organic layer was separated andconcentrated under reduced pressure at room temperature to yield thepurified product as the ammonium salt. The product was taken up in 100.0mL water and lyophilized to remove remaining traces of ammonium acetate.The lyophilized product was suspended in 40.0 mL of water, stirred with6.0 g of Chelex-100 resin [sodium form (Bio-Rad Laboratories, Hercules,Calif.], passed through a 10.0 g column of Chelex-100 resin [sodiumform], and eluted with 20.0 mL of water. The solution was filteredthrough a 0.2 μ Teflon HPLC filter (Rainin Instrument Co.) andlyophilized to provide 98.0 mg (0.063 mmol) of the tetra sodium salt,i.e., Compound 70 {R_(f) : 0.60 [chloroform:methyl alcohol:glacialacetic acid:water, 125:75:10:20v/v/v/v)]}, as a white hygroscopic foamin 87% yield.

Compound 70 is the Lipid A Analog B531-35. Lipid A Analog B531-32 wasobtained by preparing the free acid form of the analog and reacting itwith L-lysine as generally described above for ther preparation ofCompound 31 and Analog B214-32. ##STR171##

To magnetically stirred solution of 510.0 mg (0.358 mmol) of Compound 66in 6.0 mL anhydrous dichloromethane, at 0° C., was added 245.0 mg (0.895mmol) of Compound E3 (see below) and 740.0 mg (1.79 mmol) of1,3-dicyclohexylcarbodiimide. After 30 minutes, when thin layerchromatographic analysis [dichloromethane:methyl alcohol, 95:5 (v/v)]indicated completion of the reaction had occurred, the reaction mixturewas diluted with 50.0 mL ethyl acetate, filtered through 10.0 g Celite545, the solids obtained washed with 20.0 mL ethyl acetate and thefiltrate concentrated under reduced pressure at room temperature toyield a syrupy residue. The crude syrup was dissolved in 5.0 mLdichloromethane, loaded on to a silica gel (100.0 g) column and eluted,initially with a 1:3 (v/v) mixture of ethyl acetate:hexanes to removereagent residues, and then with a 2:1 (v/v) mixture of ethyl acetate:hexanes. Evaporation of solvent from the product-containing fractions(as identified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 447.0 mg (0.24 mmol) of Compound 71 {R_(f) : 0.40[dichloromethane:methyl alcohol, 95:5 (v/v)]} in 67% yield. ##STR172##

To a magnetically stirred solution of 6.0 mL 6M hydrogen fluoride inacetonitrile in a Teflon reaction vessel was added 447.0 mg (0.24 mmol)of Compound 71 dissolved in 2.3 mL of dichloromethane at roomtemperature. The resulting mixture was stirred for two hours, dilutedwith 20.0 mL saturated aqueous sodium bicarbonate solution, andextracted with 100.0 mL dichloromethane. The organic layer extract waswashed first with 20.0 mL water, and then with 10.0 mL saturated aqueoussodium chloride solution, dried over 25.0 g sodium sulfate, filtered andconcentrated under reduced pressure at room temperature. The residue waspurified on a silica gel (100.0 g) column and eluted withdichloromethane:methyl alcohol [100:4 (v/v)]. Evaporation of solventfrom the product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 404.0 mg(0.23 mmol) of Compound 72 {R_(f) : 0.37 [dichloromethane:methylalcohol, 95.5 (v/v)]} in 96% yield. ##STR173##

To a solution of 20.0 mg (0.011 mmol) of Compound 72 in 1.0 mL anhydroustetrahydrofuran, 12.5 μL (0.012 mmol) of 1.0M lithiumbis(trimethylsilyl)amide in tetrahydrofuran was added slowly under anitrogen atmosphere, at -78° C., with stirring. After five minutes, 34.0μL (0.017 mmol) of 0.5M diallyl chlorophosphate in anhydrous toluene wasadded and stirred for 10 min. The mixture was warmed to 0° C., stirred10 additional minutes, and quenched with 40.0 μL of glacial acetic acid.The reaction mixture was poured into 10.0 mL of a saturated aqueoussodium bicarbonate solution and extracted with 50.0 mL dichloromethane.The organic layer was washed first with 10.0 mL saturated aqueous sodiumbicarbonate solution and then with 10.0 mL saturated aqueous sodiumchloride solution, dried over 30.0 g sodium sulfate, filtered, andconcentrated under reduced pressure at room temperature. The residueobtained was purified on a silica gel (10.0 g) column and eluted with amixture of dichloromethan:methyl alcohol [100:4v/v)]. Evaporation ofsolvent from the product-containing fractions (as identified by thinlayer chromatographic analysis) under reduced pressure at roomtemperature and drying overnight under vacuum at room temperatureprovided 15.0 mg (0.008 mmol) of Compound 73 {R_(f) : 0.29[dichloromethane:methyl alcohol, 95.5v/v)]} in 69% yield. ##STR174##

Compound 24 (51.5 mg; 0.035 mmol) was dissolved in 1.0 mL ofdichloromethane, 0.1 mL of t-butyl alcohol and 0.1 mL of pH 7.0phosphate buffer concentrate. To this heterogeneous mixture was added100.0 mg (0.43 mmol) of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone. Themixture was magnetically stirred, under a nitrogen atmosphere, in thedark until then layer chromatographic analysis [hexanes:ethyl acetate,1:2 (v/v)] indicated complete consumption of staring material(approximately four hours). At that time, the reaction was quenched with2.0 mL 10% aqueous sodium thiosulfate solution, diluted with 10.0 mLdichoromethane and poured into 5.0 mL saturated aqueous sodiumbicarbonate solution. The organic layer was separated, washed with 5.0mL saturated aqueous sodium chloride solution, dried over 5.0 g sodiumsulfate, and filtered. The crude reaction mixture was loaded directlyonto a silica gel (10.0 g) column and eluted with dichloromethane:methylalcohol [98:2 (v/v)]. Evaporation of solvent from the product-containingfractions (as identified by thin layer chromatographic analysis) underreduced pressure at room temperature and drying overnight under vacuumat room temperature provided 40.0 mg (0.03 mmol) of Compound 74 {R_(f) :0.36 [hexane:ethyl acetate, 2:1 (v/v)]} in 87% yield. ##STR175##

To a mechanically stirred solution of 93.6 mg (0.07 mmol) of Compound 74in 1.0 mL trichloroacetonitrile, 58.0 mg (0.175 mmol) of cesiumcarbonate was added at room temperature under a nitrogen atmosphere.After one hour, the mixture was filtered through 5.0 Celite 545, thefiltered solids washed with 10.0 mL dichoromethane, and the combinedfiltrates concentrated under reduced pressure at room temperature. Thecrude product obtained was purified on a silica gel (10.0 g) column andeluted first with dichloromethane:diethyl ether [9:1v/v)] to removereagent-related impurities and then with ethyl acetate. Evaporation ofsolvent from the product-containing fractions (as identified by thinlayer chromatographic analysis) under reduced pressure at roomtemperature provided 43.2 mg (0.029 mmol) of Compound 75 {R_(f) : 0.56[dichloromethane:diethyl ether, 9:1 (v/v)]} in 42% yield. ##STR176##

To a mixture of 68.2 mg (0.046 mmol) of Compound 75 and 19.0 μL (0.092mmol) triallyl phosphite (Alfa Products) dissolved in 2.0 mL ofanhydrous dichloromethane, at 0° C., under a nitrogen atmosphere, wasadded 11.0 μL trimethylsilyl trifluoromethanesulfonate. After stirringfor one hour at 0° C., the reaction mixture was quenched with 1.0 mLsaturated sodium bicarbonate solution and extracted with 50.0 mLdichloromethane. The organic layers were dried over 25.0 g sodiumsulfate, filtered, and concentrated under reduced pressure at roomtemperature. Purification on a silica gel (25.0 g) column and elutedwith a 4:1 (v/v) mixture of hexanes: ethyl acetate and evaporation ofsolvent from the product-containing fractions (as identified by thinlayer chromatographic analysis) provided 37.0 mg (0.025 mmol) ofCompound 75 {R_(f) : 0.45 [hexanes:ethyl acetate, 2:1 (v/v)]} in 30%yield. ##STR177##

To a magnetically stirred solution of 37.0 mg (0.025 mmol) of Compound76 dissolved in 0.5 mL of anhydrous dichloromethane was added 42. 0 mg(0.076 mmol) of tin(II)tris-benzenethiolate triethylamine complex andthe resulting mixture stirred at room temperature under a nitrogenatmosphere in the dark for 30 minutes, at which time thin layerchromatographic analysis [dichloromethane:methyl alcohol, 95:5 (v/v)]showed consumption of the starting material. The reaction mixture wasloaded directly into a silica gel (5.0 g) column and eluted first with a4:1 (v/v) mixture of hexanes:ethyl acetate to remove reagentby-products, and then with 100% ethyl acetate. Evaporation of solventfrom the product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying under vacuum at room temperature for 30 minutes provided 31.9 mg(0.023 mmol) of partially purified Compound 77 {R_(f) : 0.42[dichloromethane:methyl alcohol), 95:5 (v/v)]}, which was suitable foruse in the subsequent synthetic reaction, in 90% yield. ##STR178##

To a magnetically stirred solution of 31.9 mg (0.23 mmol) of Compound 77in 0.5 mL anhydrous dichloromethane, at 0° C., was added 18.0 mg (0.92mmol) of Compound D2 (see below) and 23.0 mg (1.38 mmol) of1,3-dicyclohexylcarbodiimide. After 30 minutes, when thin layerchromatographic analysis [dichloromethane:methyl alcohol, 95:5 (v/v)]indicated completion of the reaction had occurred, the reaction mixturewas diluted with 10.0 mL ethyl acetate, filtered through 1.0 g Celite545, the solids obtained washed with 5.0 mL ethyl acetate, and thefiltrate concentrated under reduced pressure at room temperature to givea syrupy residue. The crude syrup was dissolved in 1.0 mLdichloromethane, loaded onto a silica gel (10.0 g) column and eluted,initially with a 1:4 (v/v) mixture of ethyl acetate:hexanes to removereagent residues and then with a 1:2 (v/v) mixture of ethylacetate:hexanes. Evaporation of solvent from the product-containingfractions (as identified by thin layer chromatographic analysis) underreduced pressure at room temperature and drying overnight under vacuumat room temperature provided 15.1 mg (1.04 mmol) of Compound 78 {R_(f) :0.48 [dichloromethane:methyl alcohol, 95:5 (v/v)]} in 32% yield.

To produce the Lipid A Analog B380-32, Compound 78 was deprotectedgenerally as described above for the preparation of Compound 31, and theproduct was reacted with L-lysine as described above for analog B214-32.##STR179##

To a mixture of 1.13 g (2.58 mmol) of Compound 10b and 500.0 μL (0.45mmol) allyl alcohol dissolved in 50.0 mL of anhydrous dichloromethanewas added 1.0 g of finely powdered AW-300 molecular sieves. Afterstirring one hour at room temperature, the mixture was cooled to -78°C., and 15.0 mL of a 0.02M dichloromethane solution of trimethylsilyltrifluoromethanesulfonate was added over one hour. The reaction mixturewas quenched with 10.0 mL saturated sodium bicarbonate solution andextracted with 100.0 mL dichloromethane. The organic layers were driedover 25.0 g sodium sulfate, filtered, and concentrated under reducedpressure at room temperature. Purification on a silica gel (100.0 g)column, elution with a 4:1 (v/v) mixture of hexanes:ethyl acetate, andevaporation of solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) provided 380.0 mg(0.85 mmol) of pure Compound 79b {R_(f) : 0.58 [hexanes:ethyl acetate,3:1 (v/v)]} in 33% yield and 143.0 mg (0.34 mmol) of Compound 79a {R_(f): 0.54 [hexanes:ethyl acetate, 3:1 (v/v)]} in 13% yield. ##STR180##

To a solution of 3.25 g (17.2 mmol) of Compound 79a in 220.0 mL (10:1,v/v) acetone:water was first added 6.0 g (51.2 mmol) 4-methylmorpholineN-oxide (Aldrich Chemical Co.), followed by 20.0 mg (0.08 mmol) osmiumtetroxide (Aldrich Chemical Co.). The reaction mixture was stirred fortwo and a half days at room temperature in the absence of light. Thereaction was then quenched by addition of 100.0 mL of a saturatedaqueous solution of sodium thiosulfate, stirred for an additional hour,and then extracted with 200.0 mL dichloromethane. The organic layerswere washed first with 100.0 mL water, then with 100.0 mL of a saturatedaqueous solution of sodium chloride, dried over 50.0 sodium sulfate,filtered, and concentrated under reduced pressure at room temperature togive the crude product.

The crude product was dissolved in 200.0 mL of a 1:1 (v/v) mixture ofmethyl alcohol and water. To the resulting solution was added 6.0 g(28.1 mmol) sodium periodate (Aldrich Chemical Co.), at 0° C. withvigorous stirring. After one hour, the reaction was diluted withdichloromethane. The organic layer was washed first with 100.0 mL water,then with 100.0 mL of a saturated aqueous solution of sodium chloride,dried over 50.0 g sodium sulfate, filtered, and concentrated underreduced pressure at room temperature to again give the crude product ayellowish oil.

The crude product was then dissolved in 50.0 mL methyl alcohol, cooledto 0° C. and 1.5 g(39.7 mmol) sodium bobohydride (Aldrich Chemical Co.)was added portionwise. After one hour, the reaction was quenched byaddition of 50.0 mL saturated aqueous ammonium chloride and extractedwith 200.0 mL dichloromethane. The organic layers were washed with 50.0mL saturated aqueous solution of sodium chloride, dried over 20.0 gsodium sulfate, filtered, and concentrated under reduced pressure atroom temperature to yield the crude product as a yellow brown oil.

The crude product was then dissolved in 50.0 mL methyl alcohol, and100.0 μL (25%, wt/wt) sodium methoxide was added. After two and a halfdays, the reaction was quenched by the addition of 50.0 mL saturatedaqueous ammonium chloride solution and extracted with 200.0 mLdichloromethane. The organic layers were washed with 50.0 mL saturatedaqueous solution of sodium chloride solution, dried over 50.0 g sodiumsulfate, filtered, and concentrated under reduced pressure at roomtemperature to give the crude product. The crude product was purified bycolumn chromatography on a silica gel (100.0 g), eluting with a 1:1(v/v) mixture of hexanes and ethyl acetate to provide 3.0 g (9.06 mmol)of Compound 80 {R_(f) : 0.27 [hexanes:ethyl acetate, 1:1 (v/v)]} in 53%overall yield. ##STR181##

To a solution of 1.0 g (3.02 mmol) of Compound 80 dissolved in 10.0 mLanhydrous dichloromethane, at 0° C., was added 411 mg (6.04 mmol)imidazole, followed by the addition of 0.55 g (3.62 mmol)t-butyldimethylsilyl chloride. After stirring for 30 minutes, thereaction was quenched with 10.0 mL of a saturated aqueous ammoniumchloride solution and extracted with 100.0 mL ethyl acetate. The organiclayers were dried over 50.0 g sodium sulfate, filtered, and concentratedunder reduced pressure at room temperature to give the crude product asa colorless oil. The crude product was then dissolved in 30.0 mLanhydrous dichloromethane, cooled to 0° C., and 0.97 g (3.62 mmol) of A6was added, followed by the addition of 0.75 (3.63 mmol) of1,3-dicyclohexylcarbodiimide, and 20 mg (163.7 μmol)4-dimethylaminopyridine. After two hours, the reaction mixture waswarmed to room temperature and stirred for an additional two hours,quenched with 50.0 mL saturated aqueous ammonium chloride solution, andextracted with 100.0 mL dichloromethane. The organic layers were driedover 50.0 g sodium sulfate, filtered, and concentrated under reducedpressure at room temperature to an oil. The crude oil was purified bycolumn chromatography on a silica gel (100.0 g), eluting with 1:6 (v/v)mixture of ethyl acetate and hexanes. Evaporation of the productcontaining fractions under reduced pressure at room temperature gave1.25 g (190.0 mmol) of Compound 81 {R_(f) : 0.88 [hexanes:ethyl acetate,2:1 (v/v) ]} in 63% yield. ##STR182##

To a solution of 11.2 mg (17.0 μmol) of Compound 81 dissolved in 4.0 mLof anhydrous tetrahydrofuran, at room temperature, was first added 10 μl(173 μmol) acetic acid, followed by the addition of 20 mg (76.5 μmol)solid tetrabutylammonium fluoride (Aldrich Chemical Co.). After onehour, an additional portion of 20 mg (76.5 μmol) tetrabutylammoniumfluoride was added. The reaction was stirred for one hour longer, thenquenched by the addition of 2.0 mL of a saturated aqueous ammoniumchloride solution and extracted with 50.0 mL ethyl acetate. The organiclayers were washed with 20.0 mL of a saturated aqueous sodium chloridesolution, dried over 10.0 g sodium sulfate, filtered, and concentratedunder reduced pressure at room temperature to provide 8.1 mg (14.9 μmol)of Compound 82 {R_(f) : 0.28 [hexanes:ethyl acetate, 2:1 (v/v)]} in 87%yield. ##STR183##

To a solution of 8.1 mg (14.9 μmol) of Compound 82, dissolved in 1.0 mLanhydrous dichloromethane, at 0° C., was added 7.4 mg (30.2 μmol)bis(allyloxy)(diisopropylamino) phosphine and then added 6.5 mg (92.8μmol) 1H-tetrazole. After 30 minutes, the reaction mixture was warmed toroom temperature, an additional quantity of 7.4 mg (30.2 mmol)bis(allyloxy)(diisopropylamino)phosphine was added, and the mixture wasstirred for an additional 30 minutes. The mixture was then cooled to-78° C., a solution of 6.6 mg (38.2 μmol) 3-chloroperoxybenzoic aciddissolved in 300.0 μL anhydrous dichloromethane was added, and theresulting mixture stirred for 10 minutes longer. The reaction mixturewas then quenched by the addition of 2.0 mL of a 1:1 (v/v) mixture of asaturated aqueous sodium thiosulfate solution and a saturated aqueoussodium bicarbonate solution. The resulting mixture was warmed to roomtemperature and extracted with 10.0 mL dichloromethane. The organicextracts were washed with 5.0 mL of a saturated aqueous sodium chloridesolution, and dried over 5.0 g sodium sulfate. Filtration andconcentration of the dried extract under reduced pressure at roomtemperature, gave 8.9 mg (12.6 μmol) of Compound 83 {R_(f) : 0.28[hexanes:ethyl acetate, 1:1 (v/v)]} in 85% yield. ##STR184##

A solution of 0.82 g (1.165 mmol) of Compound 83 dissolved in 100.0 mLof a 1:1 (v/v) mixture of a glacial acetic acid and water was stirred atroom temperature for eight hours. The reaction mixture was thenconcentrated, under reduced pressure at room temperature. The resultingoil was dissolved in 50.0 mL toluene and dried by azeotropic removal ofthe added toluene under reduced pressure at room temperature.

The crude oil was then dissolved in 20.0 mL anhydrous dichloromethane,cooled to 0° C., and to the oil was added 1.0 g (14.7 mmol) imidazolefollowed by 0.2 g (1.3 mmol) of tert-butyldimethylsilyl chloride. Afterstirring for 30 minutes, the reaction mixture was quenched by additionof 20.0 mL of a saturated aqueous ammonium chloride solution andextracted with 100.0 mL ethyl acetate. The organic layers were driedover 20.0 g sodium sulfate, filtered, and concentrated under reducedpressure at room temperature to yield the crude product as a colorlessoil. The crude oil was purified on a silica gel column (100.0 g) byelution with a 2:1 (v/v) mixture of hexanes:ethyl acetate. Concentrationof the product-containing fractions under reduced pressure at roomtemperature gave 0.65 g (0.835 mmol) of Compound 84 in 72% yield.##STR185##

A solution of 0.58 g (0.746 mmol) of Compound 84 dissolved in 10.0 mLanhydrous toluene was cooled to 0° C., then 1.0 mL (12.4 mmol) anhydrouspyridine, followed by 1.0 mL (1.93 mmol) of a 1.93M solution of phosgenein toluene were added. The resulting mixture was stirred for 30 minutesand then 400.0 μL (5.88 mmol) of allyl alcohol was added. After anadditional 30 minutes of stirring at room temperature, 5.0 mL of asaturated aqueous ammonium chloride solution was added. The mixture wasthen extracted with 50.0 mL dichloromethane and the organic layerswashed with 10.0 mL of a saturated aqueous sodium chloride solution,dried over 10.0 g sodium sulfate, filtered, and concentrated underreduced pressure at room temperature yielding 0.7 g (0.81 mmol) ofCompound 85 {R_(f) : 0.85 [dichloromethane:diethyl ether, 4:1 (v/v)]}.##STR186##

To a solution of 0.60 g (0.696 mmol) of Compound 85 dissolved in 5.0 mLanhydrous dichloromethane was added 0.5 g (1.1 mmol) tin(II)tris-benzenethiolate triethylamine complex. After stirring for fiveminutes, an additional 0.5 g (1.1 mmol) of tin (II)tris-benzenethiolatetriethylamine complex was added, and the mixture was stirred for anadditional five minutes. The reaction was then applied directly to ashort silica gel column (20.0 g) and eluted first with 4:1 (v/v)hexanes:ethyl acetate and then with 1:19 (v/v) methylalcohol:dichloromethane to provide the crude amine.

The crude amine was dissolved in 3.0 mL anhydrous dichloromethane,cooled to 0° C., and 290 mg (0.872 mmol) of C8 was added, followed by200.0 mg (0.969 mmol) of 1,3-dicyclohexyl carbodiimide. After one hour,the reaction mixture was allowed to warm to room temperature, stirredfor an additional two hours, diluted with 10.0 mL hexanes, filtered, andconcentrated to an oil under reduced pressure at room temperature.Purification on a silica gel column (100.0 g), by elution with a 8:1(v/v) mixture of hexanes:ethyl acetate provide 380 mg (0.330 mmol) ofCompound 86. ##STR187##

To a solution of 3.0 mL 6M hydrogen fluoride dissolved in 30.0 mLacetonitrile, contained in a Teflon reaction vessel, was added dropwiseat 0° C. a solution of 300.5 mg (0.261 mmol) of Compound 86 dissolved in2.0 mL acetonitrile. After stirring for one hour, the reaction mixturewas poured into 100.0 mL of a saturated aqueous sodium bicarbonatesolution and extracted with 100.0 mL dichloromethane. The organic layerswere washed with 50.0 mL a saturated aqueous sodium bicarbonatesolution, dried over 10.0 g sodium sulfate, filtered, and concentratedunder reduced pressure at room temperature to an oil. Purification bysilica gel column chromatography by elution with 1:1 (v/v) mixture ofhexanes:ethyl acetate provided 200 mg (0.193 mmol) of Compound 87.

Compounds 88 and 89 were synthesized by the general methods describedabove for the synthesis of Compounds 33 and 34. ##STR188##

To a stirred room temperature suspension of Compound 89 (102.5 mg, 48.6μmol) in a mixture of acetonitrile (1.5 mL) and water (100 μL) was addedmercuric oxide (96 mg, 443 μmil) and mercuric chloride (61 mg, 224μmol). After one hour, the mixture was diluted with 1:1 (v/v) methylalcohol: dichloromethane to induce precipitation. The mixture wasfiltered through Celite 545, the filtrate collected, and hydrogensulfide bubbled through it for one hour. The mixture was again filteredand the combined filtrates washed with saturated aqueous sodiumchloride, dried over sodium sulfate, filtered, and concentrated underreduced pressure to an oil. The oil was purified by a direct applicationof the oil to a silica gel column (10.0 g) and elution with 1:19 (v/v)methyl alcohol:chloroform, followed by a second silica gel column (10.0g) chromatography eluting with 1:4 (v/v) hexanes:ethyl acetate. 54.5 mg(28.3 μmol) of Compound 90 was obtained. ##STR189##

Compound 90 (37.5 mg, 19.5 μmol) was dissolved in 10:1 (v/v)tetrahydrofuran:96% formic acid under a nitrogen atmosphere in the dark,and to the solution was added tetrakis (triphenylphosphine)palladium(O)and triphenylphosphine. The reaction was carried out as generallydescribed above for Compound 30 (procedure a) and provided 15.0 mg (9.91μmol) of Compound 91 as a free acid. To produce to Lipid A AnalogB377-34, Compound 91 was reacted with L-lysine generally as describedabove for analog B214-32. ##STR190##

Compound 54 was first acylated using a standard condition of 1:1 [v/v]acetic anhydride:pyridine and a catalytic amount of4-dimethylaminopyridine added; the reaction was carried out at roomtemperature. Evaporation of the excess acylating reagents at roomtemperature under vacuum yielded the crude 4-position acylated product.This product was used in the subsequent transformation without anyfurther purification.

The crude 4-position acylated product was subjected to the synthetic andpurification steps generally described above for the transformation ofCompound 55 to 56 to provide Compound 92 {R_(f) : 0.23 [hexanes: ethylacetate, 4:1 (v/v)]}. ##STR191##

Compound 47 was methylated and treated as described above, except thattert-butylchlorodimethylsilane was used as the silylation reagent. A 2.2g (2.8 mmol) portion of isolated 4-methylated 6-silylated product wasthen dissolved in 20.0 mL of a 9:1 (v/v) mixture of acetone and water.To this mixture was added 0.7 g (5.7 mmol) 4-methylmorpholine N-oxideand 10.0 mg (0.04 mmol) osmium tetroxide, and the resulting reactionmixture was stirred for 2.5 hours. The reaction was then quenched by theaddition of 100.0. mL of saturated sodium thiosulfate solution andextracted with 100.0 mL dichloromethane. The organic layer was driedover 20.0 g sodium sulfate and evaporated under reduced pressure at roomtemperature to provide the crude product.

The crude product (obtained in the previous step) was then dissolved in20.0 mL methyl alcohol and stirred with 2.0 g potassium carbonate for 25minutes. The reaction mixture was then diluted with 100.0 mLdichloromethane, filtered thru 10.0 g Celite, and washed with 100.0 mL1.0N hydrochloric acid. The organic layer was washed with 25.0 mLsaturated sodium chloride solution, dried over 30.0 g sodium sulfate,and concentrated under reduced pressure at room temperature to yield thecrude product. The product thus obtained was purified on 200.0 g silicagel by elution with a mixture of 9:1 hexanes and ethyl acetate. Thedesired product was obtained in 80% yield {Rf: 0.46 [hexanes: ethylacetate, 4:1 (v/v)]}. The above-obtained intermediate was then subjectedto the synthetic steps generally described above for the transformationof Compound 45 to 52, followed by the synthetic steps generallydescribed above for the transformation of Compound 55 to 56 to providethe final desired intermediate, Compound 93 {R_(f) : 0.33 [hexanes:ethyl acetate, 4:1 (v/v)]}. ##STR192##

3,4,6-triacetoxygalactose (Pfanstiehl Labs., Inc.) was subjected to thesynthetic steps generally described above for the transformation ofCompound 7 to 9. The resulting product was then protected at theanomeric position as generally described above for Compound 36.Treatment of this product for the removal of the 3-,4-, and 6-acetateprotecting groups and the subsequent protection of the 4 and 6 positionswith acetonide was carried out as generally described above for thesynthesis of Compounds 36 and 5, respectively. This product was thensubjected to the synthetic steps as generally described above for thepreparation of Compounds 37 to 45, followed by the synthetic steps forthe preparation of Compounds 45 to 54.

A 10.5 g (15.6 mmol) portion of this product was dissolved in 500 mLanhydrous dichloromethane at room temperature under nitrogen, and 18.6mL (140.7 mmol) 2,4,6-collidine (Aldrich Chemical Co.) was added. Tothis mixture was next added a solution of 4.8 mL (36.3 mmol)diethylaminosulfur trifluoride (Aldrich Chemical Co.) dissolved in 120.0mL anhydrous dichloromethane over 1.5 hours dropwise. The resultingmixture was stirred 2 hours longer, then quenched by the addition of 100mL anhydrous methyl alcohol. The reaction mixture was then poured into200 mL saturated sodium bicarbonate solution and extracted with 500 mLdichloromethane. The organic extract was then washed with 200 mLsaturated aqueous sodium chloride solution, then dried over 100 g sodiumsulfate. The crude product was purified on 500 g silica gel by elutionwith a 10 to 1 (v/v) mixture of hexanes and ethyl acetate yielding thedesired 4-position fluorinated product in 65% yield {Rf: 0.77 [hexanes:ethyl acetate, 10:1 (v/v)]}.

This product was then subjected to the synthetic steps generallydescribed above for the preparation of Compound 55 to 56 to provideCompound 94 {R_(f) : 0.78 [hexanes: ethyl acetate, 2:1 (v/v)]} in goodyield. ##STR193##

Compound 95 was obtained by treatment of Compound 80 using thesilylation conditions described above for the synthesis of Compound 54,followed by alkylation with sidechain A10 (see below) using theconditions described above for the synthesis of Compound 45. Thealkylated product was then subjected to the synthetic steps generallydescribed above for the preparation of Compound 80 to 85, followed bythe synthetic steps generally described above for the preparation ofCompound 86 to 87, to provide the intermediate Compound 95 {R_(f) : 0.09[hexanes: ethyl acetate, 1:1 (v/v)]}. ##STR194##

Compound 47 was methylated and treated as described above, except thattert-butylchlorodimethylsilane was used as the silylation reagent. Theisolated 4-methylated 6-silylated product was then subjected to thesynthetic steps generally described above for the preparation ofCompound 56 from 55, followed by phosphorylation of the free 6 positionby the method described above for the synthesis of Compound 49. Thisproduct was then transformed to the desired intermediate Compound 96 (asan α β mixture) by the two step sequence described above for thesynthesis of Compounds 51A and 51B (from Compound 49 via Compound 50).Compound 96 (an α β mixture( {R_(f) : 0.50 and 0.83 [hexanes: ethylacetate, 1:1 (v/v)]}. ##STR195##

Intermediate Compound 97 was synthesized from Compound 47 in thefollowing manner. A mixture of 2.0 g (3.1 mmol) of Compound 47, 1.0 gpowdered 300AW molecular sieves, 300.0 mg (1.29 mmol)(±)-10-camphorsulfonic acid, 1.0 g (8.7 mmol) heptaldehyde and 6.0 mLanhydrous toluene were stirred for 45 minutes at room temperature undernitrogen. The reaction mixture was then diluted with 50.0 mLdichloromethane, washed first with 20.0 mL saturated aqueous sodiumbicarbonate solution, and then with 20 mL saturated aqueous sodiumchloride solution. The organic layer was dried over 30.0 g sodiumsulfate, the solvent removed under reduced pressure at room temperature,and the resulting crude product purified on 200.0 g silica gel byelution with 1.0 L hexanes followed by 1 L 99:1 (v/v) hexanes: ethylacetate and finally with 1 L 97:3 (v/v) hexanes: ethyl acetate. Thedesired product was obtained in a 85% yield (1.96 g, 2.6 mmol), R_(f) :0.47 [hexanes: ethyl acetate, 19:1 (v/v)].

The above-obtained product was dried under vacuum overnight, thendissolved in 10.0 mL anhydrous dichloromethane. To this mixture wasfirst added 625.0 μL (3.9 mmol) triethylsilane (Aldrich Chemical Co.),and next was added 2.8 mL (2.8 mmol) of a 1.0M titanium (IV) chloridedichloromethane solution (Aldrich Chemical Co.) over 5 minutes at roomtemperature under nitrogen. The reaction mixture was then diluted with50.0 mL dichloromethane, and washed first with 50 mL saturated aqueoussodium bicarbonate solution and then with 20 mL saturated aqueous sodiumchloride solution. The organic layer was dried over 10.0 g sodiumsulfate, the solvent removed under reduced pressure, at roomtemperature, and the resulting crude product purified on 200.0 g silicagel by elution with 19:1 (v/v) hexanes: ethyl acetate. The desiredproduct was obtained in a 74% yield (1.4 g, 1.9 mmol), R_(f) : 0.14[hexanes: ethyl acetate, 19:1 (v/v)].

This 6-position alkylated product was then subjected to the syntheticsteps generally described above for the three step synthetictransformation of Compounds 60A and 60B from 57A (via 58 and 59) toprovide Compound 97 (as an αβ mixture) {R_(f) : 0.55 and 0.67 [hexanes:ethyl acetate, 2:1 (v/v)]} in comparable yields. ##STR196##

Compound 12 was first reacted with Compound A25 (generally as describedabove for the synthesis of Compound 13), and the resultant product wassequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 13-23. The resultant α-isomeric productwas then reacted with Compound 17 (as generally described above for thesynthesis of Compound 24), and the resultant product was subjected tothe synthetic steps generally described above for the preparation ofCompounds 25, 32-34, 30 (Procedure b), and 31. Lipid A Analog B235-32was produced by reacting the free acid product with L-lysine asdescribed above for analog B214-32. Analog B235-31 was produced byreacting the free acid product with Tris as described above for B214-31.##STR197##

Compound 12 was reacted with Compound A6 (generally as described abovefor the synthesis of Compound 18), and the resultant product wassequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 19-23. The resultant α-isomeric productwas reacted with Compound 17 (generally as described above for thesynthesis of Compound 24), and the resultant product was sequentiallysubjected to the synthetic steps generally described above for thepreparation of Compounds 25-30 (Procedure a), with the exception thatCompound 26 was first reacted with one equivalent of an allylcarbonateprotected side chain (prepared as described below for side chains A4-A6using C4 as starting material), followed by condensation with C6. Theresultant product was then deprotected as generally described above forthe synthesis of Compound 31. Lipid A Analog B272-32 was produced byreacting the free acid product with L-lysine as described above foranalog B214-32. ##STR198##

Compound 25 was reacted with E-2-tetradecenoic acid [described in Mimuraet al., J. Pharmacobio-Dyn 1983 6(8):527, 1983] generally as describedabove for the synthesis of Compound 26, and the resultant product wassequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 27, 28 and 31. Lipid A analog B286-32was produced by reacting the free acid product with L-lysine asdescribed above for analog B214-32. ##STR199##

Compound 25 was reacted first with Compound C6 and then with Compound H1(see below) by selective condensation generally as described above forthe preparation of Compound 26, and the resultant product wassequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 27-30 (Procedure a), 31. Lipid A analogB287-32 was produced by reacting the free acid product with L-lysine asdescribed above for analog B214-32.

Compound H1 was prepared generally as described for B6 by condensingCompound C4 with Z-7-tetradecenoic acid, itself made by the same generalprocedure used to make Compound B4. ##STR200##

Compound H2 (below) was reacted with Compound 17 (generally as describedabove for the synthesis of Compound 24), and the resultant product wassequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 25-31. Lipid A analog B288-32 wasproduced by reacting the free acid product with L-lysine as describedabove for analog B214-32.

Compound H2 is identical in structure to Compound 23A, except that theallyloxy-protected phosphate group (of 23A) was replaced by anallyloxycarbonate-protected hydroxyl group (in H2); Compound H2 wasprepared essentially as described above for Compound 23A. ##STR201##

Compound 12 was reacted with Compound A17 (see below) generally asdescribed above for the preparation of Compound 18, and the resultantproduct was sequentially subjected to the synthetic steps generallydescribed above for the preparation of Compounds 19-23. The resultantα-isomeric product was then reacted with Compound 17 (generally asdescribed above for the preparation of Compound 24), and the product wassequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 25-31. Lipid A analog B294-32 wasproduced by reacting the free acid product with L-lysine as describedabove for analog B214-32. ##STR202##

Compound 12 was reacted with Compound A17 (see below) generally asdescribed above for the synthesis of Compound 18. The resultant productwas sequentially subjected to the synthetic steps generally describedabove for the preparation of Compounds 19-29 and then deprotectedgenerally as described above for Compound 31. Lipid A analog B300-32 wasproduced by reacting the free acid product with L-lysine as describedabove for analog B214-32. ##STR203##

Compound 25 was reacted first with E-2-tetradecenoic acid [described inMimura et al., J Pharmacobio-Dyn 6(8):527, 1983] and then with CompoundC5 by selective condensation (generally as described above for Compound26), and the resultant product was sequentially subjected to thesynthetic steps generally described above for the preparation ofCompounds 27, 28, and 31. Lipid A analog B313-32 was produced byreacting the free acid product with L-lysine as described above foranalog B214-32. ##STR204##

Compound 25 was reacted first with Compound H3 (by selectivecondensation) and then with Compound C6 (see below) by selectivecondensation generally as described above for the synthesis of Compound26, and the resultant product was sequentially subjected to thesynthetic steps generally described above for Compounds 27-31. Lipid Aanalog B314-32 was produced by reacting the free acid product withL-lysine as described above for analog B214-32.

Compound H3 was prepared generally as described below for Compound B6except Compound C4 was condensed with decanoic acid (Aldrich Chm. Co.).##STR205##

Compound 25 was reacted with a racemic mixture of E3 and E5 generally asdescribed above for the synthesis of Compound 26, and the resultantproduct was sequentially subjected to the synthetic steps generallydescribed above for the preparation of Compounds 27, 28, 31. Lipid Aanalog B318-32 was produced by reacting the free acid product withL-lysine as described above for analog B214-32. ##STR206##

Compound 23A was reacted with Compound 87 generally as described abovefor the synthesis of Compound 24. The resultant product was thensubjected first to the synthetic steps generally described above for thepreparation of Compound 32-34, 30 (Procedure b), and 31 (in that order)and then deprotected generally as described above for Compound 31. LipidA analog B377-34 was prepared by reacting the free acid product withL-lysine generally as described above for analog B214-32, except that adilysine (rather than a tetralysine) salt is produced. ##STR207##

Compound 12 was reacted with Compound A23 (see below) generally asdescribed above for the synthesis of Compound 18, and the resultantproduct was sequentially subjected to the synthetic steps generallydescribed above for the preparation of Compound 19-23. The resultantα-isomeric product was then reacted with Compound 17 generally asdescribed above for the synthesis of Compound 24, and the product wassubjected to the synthetic steps generally described above for thepreparation of Compounds 32-34, 30 (Procedure b), and 31 (in thatorder). Lipid A analog B379-32 was produced by reacting the free acidproduct with L-lysine as described above for analog B214-32. ##STR208##

Compound 25 was reacted with Compound A30 (below) generally as describedabove for Compound 26. The resultant product was first sequentiallysubjected to the synthetic steps generally described above for thepreparation of Compounds 27 and 28 and then deprotected as describedabove for the preparation of Compound 31. Lipid A analog B385-32 wasproduced by reacting the free acid product with L-lysine as describedabove for analog B214-32. ##STR209##

Compound 25 was first reacted with Compound A31 (below) generally asdescribed above for Compound 26. The resultant product was firstsequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 27 and 28 and then deprotected asdescribed above for the preparation of Compound 31. Lipid A analogB387-32 was produced by reacting the free acid product with L-lysine asdescribed above for analog B214-32. ##STR210##

Compound 25 was first reacted with Compound C8 and then reacted withE-2-tetradecenoic acid [described in Mimura et al., J. Pharmacobio-Dyn6(8): 527, 1983] generally as described above for the synthesis ofCompound 26. The resultant product was sequentially subjected to thesynthetic steps generally described above for the preparation ofCompounds 27, 28, 30 (Procedure b), and 31. Lipid A analog B388-32 wasproduced by reacting the free acid product with L-lysine as describedabove for analog B214-32. ##STR211##

Compound 25 was reacted with Compound G2 generally as described abovefor the synthesis of Compound 32. The resultant product was firstsequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 33 and 34, and then the oxythiolanegroups were deprotected by the method described for the preparation ofCompound 30 (Procedure b) above, and the phosphate and hydroxyl groupswere deprotected by the method described for Compound 31 above. Lipid Aanalog B398-32 was produced by reacting the free acid product withL-lysine as described above for analog B214-32. ##STR212##

Compound 12 was first reacted with trans-2-decenoic acid (LancasterSynthesis Inc.) generally as described above for the synthesis ofCompound 18, and the resultant product sequentially subjected to thesynthetic steps generally described above for the preparation ofCompounds 19-23. The resultant α-isomeric product was next reacted withCompound 17 (generally as described above for the preparation ofCompound 24), and the product subjected to the synthetic step generallydescribed above for the preparation of Compound 25. The product obtainedwas then reacted generally as described above for the synthesis ofCompound 32, the resultant product subjected to the synthetic stepsgenerally described above for the preparation of Compounds 33, 34, and30 (Procedure b) (in that order), and the phosphate and hydroxyl groupsdeprotected generally as described above for the preparation of Compound31. Lipid A analog B400-32 was produced by reacting the free acidproduct with L-lysine as described above for analog B214-32. ##STR213##

Compound 12 was reacted with Compound A6 (generally as described abovefor the synthesis of Compound 18), and the resultant product wassequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 19-23. The α-isomeric product wastermed H12A.

Compound 12 was reacted with H1 (see preparation of Compound B287)generally as described above for the synthesis of Compound 13, and theproduct subjected first to the synthetic steps generally described abovefor the preparation of Compound 19, and then to the synthetic stepsgenerally described above for the preparation of Compounds 15-17 (inthat order). The product was termed H13.

H12A was reacted with H13 generally as described for the preparation ofCompound 24, and the product was subjected first to the synthetic stepgenerally described above for the preparation of Compound 25, then tothe synthetic steps generally described above for the preparation ofCompounds 32-34 (in that order). Finally, the dithiane groups of theproduct were deprotected as generally described above for Compound 30(Procedure b), and the phosphate and hydroxyl groups were deprotected asgenerally described above for the synthesis of Compound 31. Lipid AAnalog B406-32 was produced by reacting the free acid product withL-lysine as described above for analog B214-32. ##STR214##

Compound 53 was first reacted with Compound 23A (generally as describedabove for the synthesis of Compound 40), and the resultant productsequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 41-44. The product obtained was thenphosphorylated as generally described above for the preparation ofCompound 30 (Procedure b), and the phosphate and hydroxyl groupsdeprotected generally as described for the preparation of Compound 31.Lipid A analog B410-32 was produced by reacting the free acid productwith L-lysine as described above for analog B214-32. ##STR215##

Compound 51A was first reacted with Compound 39 (generally as describedabove for the synthesis of Compound 40), and the resultant productsequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 41-44. The product obtained was thenphosphorylated as generally described above for the preparation ofCompound 30 (Procedure b), and the phosphate and hydroxyl groupsdeprotected generally as described for the preparation of Compound 31.Lipid A analog B415-32 was produced by reacting the free acid productwith L-lysine as described above for analog B214-32. ##STR216##

Compound 42 was reacted with 1,3-dicyclohexylcarbodiimide and E7generally as described above for the preparation of Compound 43, and theresultant product first subjected to the synthetic step generallydescribed above for the preparation of Compound 44, then phosphorylatedas generally described above for the preparation of Compound 30(Procedure b), and finally deprotected as generally described above forthe synthesis of Compound 31. The analog B425-32 was then produced byreacting the free acid product with L-lysine as described above foranalog B214-32. ##STR217##

A minor anomeric glycosidation product of the synthetic reactionproducing Compound 40 (above) was first sequentially subjected to thesynthetic steps generally described above for the preparation ofCompounds 41-44, phosphorylated as generally described above forCompound 30 (Procedure c), and deprotected as generally described abovefor Compound 31. The analog B426-32 was then produced by reacting thefree acid product with L-lysine as described above for analog B214-32.##STR218##

Compound 51A was first reacted with Compound 56 (generally as describedabove for the synthesis of Compound 65), and the resultant productsequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 66-69. The product obtained was thendeprotected generally as described for the preparation of Compound 31.Lipid A analog B427-32 was produced by reacting the free acid productwith L-lysine as described above for analog B214-32. ##STR219##

Compound 56 was first reacted with Compound 23A (generally as describedabove for the synthesis of Compound 24), and the resultant productsubjected to the synthetic step generally described above for thepreparation of Compound 25. The resultant product was then reacted witha mixture of Compounds E3 and E5 (see below) generally as describedabove for Compound 43. The product was then subjected to the syntheticsteps generally described above for the preparation of Compounds 33,phosphorylated as generally described above for the preparation ofCompound 28, and the product deprotected generally as described for thepreparation of Compound 31. Lipid A analog B422-32 was produced byreacting the free acid product with L-lysine as described above foranalog B214-32. ##STR220##

Compound 19 was subjected to the same synthetic steps as for thesynthesis of Compounds 58 from Compound 47. The resulting product wasdeprotected as generally described above for Compound 22 and thenactivated as generally described above for the synthesis of Compound23A. This product was first reacted with Compound 56 (generally asdescribed above for the synthesis of Compound 65), and the resultantproduct subjected to the synthetic step generally described above forthe preparation of Compound 66. The product was then reacted withCompound E3 (see below) generally as described for the preparation ofCompound 67 and the product subjected to the synthetic steps generallydescribed above for the preparation of Compounds 68, 69, 31 (in thatorder). Lipid A analog B451-32 was produced by reacting the free acidproduct with L-lysine as described above for analog B214-32. ##STR221##

Compound 25 was first reacted with Compound E3 (generally as describedabove for the synthesis of Compound 26), and the resultant productsubjected to the synthetic steps generally described above for thepreparation of Compounds 27, 28, 31 (in that order). Lipid A analogB452-32 was produced by reacting the free acid product with L-lysine asdescribed above for analog B214-32. ##STR222##

Compound 51A was first reacted with Compound 56 (generally as describedabove for the synthesis of Compound 65), and the resultant productsubjected to the synthetic step generally described above for thepreparation of Compound 66. The product was then reacted with a mixtureof Compounds E3 and E5 as generally described above for the synthesis ofCompound 67, and the product subjected to the synthetic steps generallydescribed for the preparation of Compounds 68, 69, 31 (in that order).Lipid A analog B459-32 was produced by reacting the free acid productwith L-lysine as described above for analog B214-32. ##STR223##

Compound 51A was first reacted with Compound 56 (as generally describedabove for the synthesis of Compound 65), and the resultant productsubjected to the synthetic step generally described above for thepreparation of Compound 66. The resultant product was then reacted withCompound E3 (see below) as generally described for the synthesis ofCompound 67, and the product sequentially subjected to the syntheticsteps generally described above for the preparation of Compounds 68 and69. The product obtained was then deprotected generally as described forthe preparation of Compound 31. Lipid A analog B460-32 was produced byreacting the free acid product with L-lysine as described above foranalog B214-32. ##STR224##

Compound 51A was first reacted with Compound 56 (as generally describedabove for the synthesis of Compound 62). and the resultant productsubjected to the synthetic step generally described above for thepreparation of Compound 66. The resultant product was then reacted withCompound E5 (see below) as generally described for the synthesis ofCompound 67, and the product sequentially subjected to the syntheticsteps generally described above for the preparation of Compounds 68 and69. The product obtained was then deprotected generally as described forthe preparation of Compound 31. Lipid A analog B465-32 was produced byreacting the free acid product with L-lysine as described above foranalog B214-32. ##STR225##

A mixture of Compounds 60A and 60B was first reacted with Compound 56(as generally described above for the synthesis of Compound 65), and theresultant product subjected to the synthetic step generally describedabove for Compound 66. The resultant product was then reacted withCompound E3 (see below) as generally described for the synthesis ofCompound 67, and the product subjected to the synthetic steps generallydescribed above for the preparation of Compounds 68, 69, and 31 (in thatorder). Lipid A analog B466-32 was produced by reacting the free acidproduct with L-lysine as described above for analog B214-32. ##STR226##

A mixture of Compounds 60A and 60B was first reacted with Compound 56(generally as described above for the synthesis of Compound 65), and theresultant product subjected to the synthetic step generally describedabove for the preparation of Compound 66. The product was then reactedwith Compound E5 as generally described above for the synthesis ofCompound 67, and the product subjected to the synthetic steps generallydescribed for the preparation of Compounds 68, 69, and 31 (in thatorder). Lipid A analog B477-32 was produced by reacting the free acidproduct with L-lysine as described above for analog B214-32. ##STR227##

A mixture of Compounds 60A and 60B was first reacted with Compound 56(generally as described above for the synthesis of Compound 65), and theresultant product sequentially subjected to the synthetic stepsgenerally described above for the preparation of Compounds 66-68. Theproduct (corresponding to Compound 68 above) was then deprotectedgenerally as described for the synthesis of Compound 31. Lipid A analogV479-33 was produced by reacting the free acid product with L-lysine asdescribed above for analog B214-32. ##STR228##

Compound 52 was first sequentially subjected to the reactions generallydescribed above for the preparation of Compounds 46 and 47 and thensequentially subjected to the reactions generally described above forthe preparation of Compounds 57-60. The resultant product was thenreacted with Compound 56 (as generally described above for the synthesisof Compound 65), and the resultant product was sequentially subjected tothe synthetic steps as generally described above for the preparation ofCompounds 66-70 to produce Analog B510-35. ##STR229##

Analog B464 is identical in structure to Compound 70 with the exceptionthat in the preparation of Compound 45 a one carbon extended sidechainanalog of A10 was used (the preparation of A10 was modified by the useof octyl cyanide). ##STR230##

Compound 92 was coupled with Compound 60 (as generally described abovefor the synthesis of Compound 65). The resultant product was thensequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 66-70 to produce Analog B718-35.##STR231##

Compound 93 was coupled with Compound 60 (as generally described abovefor the synthesis of Compound 65). The resultant product was thensequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 66-70 to produce Analog B587-35.##STR232##

Compound 94 was coupled with Compound 60 (as generally described abovefor the synthesis of Compound 65). The resultant product was thensequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 66-70 to produce Analog B737-35.##STR233##

Compound 95 was coupled with Compound 60 (as generally described abovefor the synthesis of Compound 65). The resultant product was thensequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 66-70 to produce Analog B736-35.##STR234##

Compound 96 was coupled with Compound 56 (as generally described abovefor the synthesis of Compound 65). The resultant product was thensequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 66-70 to produce Analog B725-35.##STR235##

Compound 97 was coupled with Compound 56 (as generally described abovefor the synthesis of Compound 65). The resultant product was thensequentially subjected to the synthetic steps generally described abovefor the preparation of Compounds 66-70 to produce Analog B763-35.##STR236##

To refluxing anhydrous tetrahydrofuran (500.0 mL) was sequentiallyadded: activated zinc (101.0 g, 1.54 mol; Fisher Scientific), ethylbromoacetate (3.0 mL; Aldrich Chemical Co.), and , in one portion,heptyl cyanide (47.4 mL, 0.308 mol; Aldrich Chemical Co.). To theresulting mixture, 134.0 mL (1.232 mol) of ethyl bromoacetate was thenadded dropwise, over three hours. The mixture was refluxed for anadditional 10 min, cooled to room temperature, and quenched by the slowaddition of saturated aqueous potassium carbonate solution (160.0 mL).Following rapid stirring for 30 minutes, the solution was filteredthrough 550.0 g Celite 545 to yield a clear yellow solution of crudeenamino ester. The solution was acidified with 1.0N hydrochloric acid(300.0 mL), stirred for three hours, diluted with 2.0 L hexanes, andneutralized by the addition of 300.0 mL saturated aqueous sodiumhydrogen carbonate. The organic layer was washed with saturated aqueoussodium chloride solution (440.0 mL), dried over 500.0 g sodium sulfate,filtered, and evaporated. The residue was purified by application to asilica gel (1.0 kg) and elution with 6:1 (v/v) hexanes/ethyl acetate.Evaporation of solvent from the product-containing fractions (asdetermined by thin layer chromatographic analysis) under reducedpressure at room temperature provided 64.0 g (0.298 mol) of Compound A1.{R_(f) : 0.7 [hexanes:ethyl acetate, 4:1 (v/v)]} in 97% yield.##STR237##

[R]-(+)-2,2'-Bis(diphenylphosphino)-1,1'-binaphthyl (653.5 mg, 1.05mmol; Aldrich Chemical Co.) and cyclooctadienyl ruthenium dichloride(279.8 mg, 1.0 mmol; Alfa Chemical Co., Ward Hill, Mass.) were combinedin a 125 mL stopcock sidearm round bottom flask fitted with a magneticstirrer and cold finger water condenser in a dry box. The flask wasremoved from the dry box and placed under argon. Anhydrous toluene (40.0mL) and triethylamine (1.7 mL, 10.0 mmol; Aldrich Chemical Co.), both ofwhich had been deoxygenated by sparging with nitrogen, were injectedinto the flask, and the mixture was refluxed under argon, with stirring,for 15 hours. The deep crimson solution was allowed to cool to 20° C.and formed a reddish gel. Excess solvent was removed from the mixtureusing a 12-inch, 22-gauge needle, and the residual volatiles wereremoved by application of a vacuum, over several hours (using great careto exclude air). The residual red-black solid was dissolved inanhydrous, oxygen-free tetrahydrofuran by stirring under a nitrogenatmosphere at 25° C. for one hour. The resultant clear orange-brownsolution of [R]-2,2'-Bis(diphenylphosphino)-1,1'-binaphthyl rutheniumdichloride hemi triethylamine complex was used directly in the nextreaction.

Compound A1 (334.2 g, 1.15 mol) was dissolved in anhydrous methylalcohol (330.0 mL) and was deoxygenated by three freeze-thaw vacuumdegassing cycles, using liquid nitrogen and a nitrogen atmosphere. Thesolution of [R]-2,2'-Bis(diphenylphosphino)-1,1'-binaphthyl rutheniumdichloride hemi triethylamine complex catalyst (prepared above), wasadded to the reaction solution using a syringe. The reaction mixture waspumped into an argon-flushed 2.0 L hydrogenation bomb containing methylalcohol-washed Dowex 50×8-200 H⁺ resin (3.0 g; Aldrich Chemical Co.)using a catheter under argon. The bomb was charged to 1480 psi withhydrogen gas (Liquid Carbonic, Tewskbury, Mass.) and the reactionmixture was stirred at 25° C. for 66 hours. When the pressure in thebomb had dropped 300 psi, excess hydrogen gas was released, the reactionmixture was filtered, and volatile substances were removed, underreduced pressure, to provide 334.0 g (1.15 mol) of Compound A2 {R_(f) :0.31 [hexanes:ethyl acetate, 4:1 (v/v)]} in 99% yield. ##STR238##

Compound A2 (89.6 g, 0.347 mol) was dissolved in tetrahydrofuran (800.0mL). To this solution was added 2.5M aqueous sodium hydroxide (300.0 mL,0.75 mol), and the resulting mixture vigorously stirred, under anitrogen atmosphere, at 25° C., for one and a half hours. The reactionmixture was diluted with 1.0 L of 1:1 (v/v) diethyl ether/hexanes, andthe aqueous layer was separated. The organic phase was extracted furtherwith 200.0 mL of water, and the combined aqueous phases acidified withconcentrated hydrochloric acid 67 mL. The acidified mixture was thenextracted with 2.0 L diethyl ether and the extract washed first with 1.0L water, then with 1.0 L saturated aqueous sodium chloride solution, andfinally dried over 500.0 g magnesium sulfate. The solvent was removedunder reduced pressure, and the resultant grayish solid dissolved in 2.0L 80° C. acetonitrile. To the solution at 80° C. was addeddicyclohexylamine (80.0 mL, 0.40 mol; Aldrich Chemical Co.). The mixturewas cooled to -20° C., providing 104.7 g (0.24 mol) of Compound A3{R_(f) : 0.38 [hexanes:ethyl acetate:glacial acetic acid, 1:1:0.1(v/v/v)]} as slightly off-white fine needles, in 71% yield. ##STR239##

Compound A3 (104.7 g, 246.0 mmol) was suspended in ethyl acetate (2.0L), and, to the suspension, triethylamine (37.2 g, 369.0 mmol) was firstadded, followed by 2-bromoacetophenone (48.9 g, 246.0 mmol), in oneportion; Aldrich Chemical Co.); additions were made under a nitrogenatmosphere, at 0° C. After three hours, the reaction mixture was warmedto room temperature, stirred for six hours, and then vacuum filtered.The residue was washed with 400.0 mL ethyl acetate, and the filtrate waswashed first with 500.0 mL of 0.8M hydrochloric acid, then with 500 mLof water, and finally with 1.0 mL saturated aqueous sodium chloridesolution, and then dried over 500.0 g magnesium sulfate. The solvent wasevaporated under reduced pressure at 50° C. to yield a congealed graysolid which was recrystallized from 1:1 L hexanes and dried in a vacuumoven at 50° C. to provide 81.05 g (223.9 mmol) of Compound A4 {R_(f) :0.65 [chloroform:methyl alcohol, 95:5 (v/v)]} as off-white solid in 91%yield. ##STR240##

Compound A4 (20.2 g, 65.9 mmol) was dissolved in anhydrous toluene(300.0 mL) and anhydrous pyridine (30.0 mL) at 0° C. and, to thissolution, was added 1.93M phosgene in toluene (50.0 mL, 96.5 mmol),dropwise. The reaction mixture was stirred for 10 minutes, and thenallyl alcohol (20.2 mL, 297.0 mmol) was added dropwise. After anadditional 10 minutes of stirring, the reaction was quenched, at 0° C.,by the addition of 100.0 mL saturated sodium bicarbonate solution. Thesolution was subsequently warmed to 25° C. and extracted with 1.0 Lethyl acetate. The organic layer was washed with 500.0 mL saturatedaqueous sodium chloride solution, dried over 500.0 g sodium sulfate,filtered, and evaporated. The residue was purified on a silica gel (2.0kg) column, eluting with 1:9 (v/v) ethyl acetate/hexanes. Evaporation ofsolvent from the product-containing fractions (as determined by thinlayer chromatographic analysis) under reduced pressure at roomtemperature provided 16.1 g (41.2 mmol) of Compound A5 {R_(f) : 0.9[hexanes:ethyl acetate, 2:1 (v/v)]} in 62% yield. ##STR241##

Compound A5 (16.07 g, 41.17 mmol) was dissolved in glacial acetic acid(150.0 mL), in a Morton flask, at 0° C. and, to the solution, was addedzinc dust (24.2 g, 371.0 mmol). The solution was warmed to 25° C.,stirred for one hour, and then filtered through a 50.0 g Celite 545 plugand evaporated. The residue was purified on a silica gel column, elutingfirst with 4:1 (v/v) ethyl acetate/hexanes and then with 10:40:1 (v/v/v)methyl alcohol/chloroform/acetic acid. Evaporation of solvent from theproduct-containing fractions (as determined by thin layerchromatographic analysis) under reduced pressure at room temperatureprovided A6 {10.8 g; 39.65 mmol; R_(f) : 0.34 [hexanes:ethyl acetate,2:1 (v/v)]} in 96% yield. ##STR242##

To a mechanically stirred suspension of 1003.0 g (15.33 mol) ofactivated zinc powder in 2.5 L of anhydrous tetrahydrofuran under anitrogen atmosphere at room temperature was added dropwise 56.0 mL (0.59mol) of methyl bromoacetate (Lancaster Chemical Co., Windham, N.H.) overa 10-minute period. The reaction mixture was warmed to refluxtemperature and 496.3 g (3.96 mol) of n-heptyl cyanide (Aldirch ChemicalCo.) was added dropwise over a five-minute period, and an additional700.0 mL (7.39 mol) of methyl bromoacetate was then added (dropwise)over a four-hour period. The mixture was refluxed for one additionalhour, allowed to cool to room temperature, slowly poured into 3.0 L of astirred, saturated aqueous solution of potassium carbonate, and 1.0 kgof Celite 545 was added. The heterogeneous mixture was filtered over apad of 200.0 g of Celite 545 and eluted with four 1.0 L portions ofethyl acetate. The filtrate was separated, and the aqueous layerextracted with two 500.0 mL portions of ethyl acetate. The combinedorganic layers were washed with 500.0 mL of a saturated aqueous solutionof sodium chloride, dried over 2.0 kg of sodium sulfate, filtered, andconcentrated under reduced pressure, at room temperature. The crudeorange oil was vigorously stirred at room temperature in a two phasesystem of 1.5 L of hexanes and 500.0 mL of a 1.0N aqueous solution ofhydrochloric acid with a dropwise addition of 250 mL concentratedhydrochloric acid over a forty minute period. After stirring the finalheterogeneous solution for an additional 20 minutes, the layers wereseparated and the aqueous layer was extracted with two 200.0 mL portionsof hexanes. The combined organic layers were washed with 500.0 mL of asaturated aqueous solution of sodium hydrogen carbonate, dried over500.0 g of sodium sulfate, filtered, and concentrated under reducedpressure, at room temperature. The crude orange liquid was distilledusing a brush rotary distillation apparatus at a bath temperature of110° C., under 1.0 mm Hg vacuum. The partially purified clear yellow oilwas fractionally vacuum-distilled to provide 652.8 g [3.26 mol, 82.3%boiling point (b.p.) 86°-88° C./0.4 mm Hg) of Compound A7 {R_(f) : 0.65[hexanes:ethyl acetate, 4:1 (v/v)]} as a clear, colorless liquid.##STR243##

In an oxygen-free dry box, 1.54 g (2.47 mmol) of[R]-(+)-2.2'-bis(diphenylphosphino)-1,1'-binaphthyl and 662.0 mg (2.36mmol) dichloro(cycloocta-1,5-diene)ruthenium (II) polymer were suspendedin 100.0 mL of degassed toluene and 4.0 mL oxygen-free triethylamine ina 250 mL Schlenk flask equipped with a magnetic stirring bar and acondenser. The reaction vessel was sealed under the inert atmosphere,removed from the dry box, and refluxed under an argon atmosphere untilan orange solid was obtained (approximately 24 hours). The reactionmixture was cooled slowly to 0° C. over a two-hour period, after whichtime the gelatinous red semisolid was suspended in 50.0 mL of dry,degassed toluene. The suspension was lightly swirled to wash thecrystaline-like sheets, allowed to stand 10 minutes, and the excesssolvent was decanted from the solid using a 50 mL syringe with a20-gauge needle. The above trituration was repeated one additional time,and this was followed by evaporating the final catalyst to dryness undera 1 mm Hg vacuum over a two-hour period. The orange-red solid wassuspended in 100.0 mL anhydrous, oxygen-free tetrahydrofuran and stirredunder an argon atmosphere for one hour during which time the mixturebecame a clear red solution. This catalyst solution was transferred viaa canula, under an argon atmosphere, to the degassed Compound A7solution described below.

Compound A7 (365.3 g; 1.824 mol) was dissolved in 500.0 mL of a freshlyopened bottle of HPLC grade methyl alcohol, under an argon atmosphere,in a 2 L three-necked round bottom flask. The flask was sealed with avacuum adapter and two rubber septa, and the solution was cooled, withliquid nitrogen, to a white solid during which time the flask wasevacuated under reduced pressure. The solid was then placed under anargon atmosphere and warmed to room temperature with the aid of a heatgun. Such a cooling, evacuation, and warming process was repeated threeadditional times. After the final degassing process, the chiral catalystin 100.0 mL of anhydrous, oxygen-free tetrahydrofuran was added asdescribed above. The final reaction mixture was transferred using aTeflon canula under an argon atmosphere into a 2.0 L reaction bomb whichcontained 1.0 g (5.21 mmol) of para-toluenesulfonic acid monohydrate(Aldrich Chemical Co.) and which had been purged with argon for twohours. (The reaction bomb was equipped with a mechanical stirrer andpressure gauge.) The reaction bomb was evacuated using water aspirationand purged twice with 100 psi of hydrogen gas. The reaction waspressurized to 1500 psi with hydrogen gas and allowed to stir for 72hours; the system was repressurized after the first 15 minutes ofstirring. After the loss of 360 psi of hydrogen gas, the completedreaction was slowly depressurized and purged three times with argon. Themethanolic solution was evaporated under reduced pressure, and theresulting residue was dissolved in ethyl acetate and stirred with 300.0mL of a saturated aqueous solution of sodium bicarbonate for 15 minutes.The layers were separated, and the organic layer was washed with 100.0mL of a saturated aqueous solution of sodium chloride, dried over 100.0g of sodium sulfate, filtered, and concentrated under reduced pressure,at room temperature. The residue was purified over 2.5 kg of silica geleluting first with 32.0 L of hexanes, then with 8.0 L of 19:1 ((v/v)hexanes:ethyl acetate, then with 16.0 L of 9:1 (v/v) hexanes:ethylacetate, and finally with 8.0 L of 3:1 (v/v) hexanes:ethyl acetate.Evaporation of the solvent from the-containing fractions (as identifiedby thin layer chromatographic analysis) under reduced pressure, at roomtemperature, and drying overnight, under vacuum, at room temperatureprovided 325.0 g (1.61 mol, 88.1% yield, 98+% enantiomeric excess) ofCompound A8 {R_(f) : 0.46 [hexanes:ethyl acetate, 3:1 (v/v)]} as a clearand colorless oil. ##STR244##

To a stirred suspension of 31.5 g (0.83 mol) of lithium aluminum hydride(Aldrich) in 500.0 g of anhydrous diethyl ether, under a nitrogenatmosphere, at 0° C., was added dropwise 159.0 g (0.78 mol) of CompoundA8 in 200.0 g anhydrous diethyl ether over a three and a half-hourperiod. After stirring for an additional 15 minutes at room temperature,the completed reaction was cooled to 0° C. and quenched with a dropwiseaddition of 1.0 L of a 2.0N aqueous solution of hydrochloric acid,followed by addition of 200.0 mL of concentrated hydrochloric acid. Theresulting clear layers were separated, and the aqueous layer wasextracted three times with 300.0 mL portions of diethyl ether. Thecombined extracts were washed first with 200.0 mL of water and then with200.0 mL of a saturated aqueous solution of sodium chloride. The aqueouslayers were back extracted three times with 300.0 mL portions ofchloroform. The combined organic layers were dried over 500.0 g ofsodium sulfate, filtered, and concentrated under reduced pressure atroom temperature to provide a clear yellow oil. The crude product waspurified on 500.0 g of silica gel eluting first with 5.0 L of 9:1 (v/v)hexanes:ethyl acetate, then with 20.0 L of 4:1 (v/v) hexanes:ethylacetate, then with 8.0 L of 3:1 (v/v) hexanes:ethyl acetate, then with1.0 L of chloroform, then with 6.0 L of 9:1 (v/v) chloroform:methylalcohol, and finally with 4.0 L of 4:1 (v/v) chloroform:methyl alcohol.Evaporation of the solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 94.2 g (0.54 mol, 69% yield) of Compound A9 {R_(f): 0.33 [ethyl acetate:hexanes, 1:1 (v/v)]} as a clear colorless oil.##STR245##

To a stirred solution of 114.1 g (0.65 mol) of Compound A9 in 3.6 L ofanhydrous pyridine, at 2.0° C., under a nitrogen atmosphere, was added136.6 g (0.72 mol) of para-toluenesulfonyl chloride (99+%, AldrichChemical Co.) in 10.0 g portions over a 15-minute period. The reactionwas allowed to warm slowly to room temperature, was stirred for eighthours under a nitrogen atmosphere, was concentrated under vacuumevaporating conditions, and azeotroped to dryness with three 500.0 mLportions of toluene using vacuum evaporation. The crude syrup wasdissolved in 2.5 L of ethyl acetate and 500.0 mL of a saturated aqueoussolution of sodium chloride. The layers were separated and the organiclayer was washed with 500.0 mL of a saturated aqueous sodium chloridesolution. The combined aqueous layers were extracted twice with 500.0 mLportions of chloroform. The combined organic layers were dried over500.0 of sodium sulfate, filtered and concentrated under reducedpressure, at room temperature. The residue was purified over 1.5 kg ofsilica gel eluting first with 12.0 L of 9:1 (v/v) hexanes:ethyl acetate,then with 12.0 L of 17:3 (v/v) hexanes:ethyl acetate, then with 20.0 Lof 4:1 (v/v) hexanes:ethyl acetate, then with 4.0 L of dichloromethane,and finally with 16.0 L of 9:1 (v/v) dichloromethane:methyl alcohol.Evaporation of the solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 96.9 g (0.29 mol) of Compound A10 {R_(f) : 0.45[hexanes:ethyl acetate, 2:1 (v/v)]} as a yellow oil in 45% yield.##STR246##

To a stirred solution of 20.0 mL (0.136 mol) of 1-octyne (AldrichChemical Co.) in 300.0 mL of anhydrous tetrahydrofuran was addeddropwise 70.5 mL (0.177 mol) of a 2.5M solution of n-butyl lithium inhexanes (Aldrich chemical Co.), under a nitrogen atmosphere, at 0° C.,over a 30-minute period. The mixture was allowed to warm to roomtemperature over a one hour period, after which time the reactionmixture was cooled to 0° C. and 35.9 mL (0.272 mol) of 1,4-diiodobutane(Aldrich Chemical Co.) was added dropwise over a 20-minute period. Themixture was allowed to warm to room temperature, stirred for anadditional 16 hours, diluted with 300.0 mL hexanes, poured over 400 g ofice, and the resulting layers separated. The organic layer was washedwith 300.0 mL a saturated aqueous solution of sodium chloride, driedover 150.0 g of sodium sulfate, filtered, and concentrated under reducedpressure at room temperature. The crude product was rendered free ofexcess diiodobutane by distillation under 0.1 mm Hg of vacuum at 70°-80°C., and the remaining residue was purified on 500.0 g of silica gel byeluting with 2.0 L of hexanes. Evaporation of the solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure, at room temperature,and drying overnight, under vacuum, at room temperature provided 23.0 g(0.078 mol) of Compound A11 {R_(f) : 0.6 [hexanes]} in 58% yield.##STR247##

To a stirred solution of 6.53 g (37.5 mmol) of Compound A9 in 45.0 mL ofanhydrous pyridine was added 11:58 g (37.5 mmol) of4-methoxy-triphenylmethyl chloride (Aldrich Chemical Co.), under anitrogen atmosphere at room temperature. The reaction mixture wasstirred at room temperature for four and a half hours, diluted with200.0 mL of dichloromethane, and the organic solution washed with 100.0mL water, dried over 150.0 g of sodium sulfate, filtered, concentratedunder reduced pressure at room temperature, and azeotroped to drynesswith three 100 mL portions of toluene under vacuum evaporation. Thecrude product was purified on 300.0 g of silica gel eluting with 2.0 Lof 6:1 (v/v) hexanes:ethyl acetate. After evaporation of the solventfrom the product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature theproduct was dissolved in 200 mL of hexanes, filtered, and the filtratewas concentrated under reduced pressure at room temperature and driedovernight under vacuum at room temperature to provide 16.5 g (36.9 mmol)of Compound A12 {R_(f) : 0.49 [hexanes:ethyl acetate, 4:1 (v/v)]} in 98%yield. ##STR248##

to a stirred solution of 17.1 g (0.038 mol) of Compound A12 in 60.0 mLanhydrous N,N-dimethylformamide, under a nitrogen atmosphere, at 0° C.,was added (in small portions) 2.94 g (0.076 mol) of sodium hydride (60%in oil, Aldrich chemical Co., washed with hexanes). The mixture wasstirred at 0° C. for an additional 15 minutes, 12.3 g (0.042 mol) ofCompound A11 was added dropwise over a 30minute period, the reactionmixture was allowed to warm to room temperature, stirred for anadditional 16 hours, and quenched with the slow addition of 10.0 mL ofmethyl alcohol at 0° C. The mixture was stirred for an additional 30minutes, diluted with 300.0 mL dichloromethane, and the resultingorganic solution washed with 200.0 mL of a saturated aqueous solution ofsodium chloride, dried over 150.0 g of sodium sulfate, filtered, andconcentrated under reduced pressure, at room temperature. The residuewas purified on 500.0 g of silica gel eluting first with 1.5 L ofhexanes and then with 2.5 L of 30:1 (v/v) hexanes:ethyl acetate.Evaporation of the solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 2.5 g (4.1 mmol) of Compound A13 {R_(f) : 0.5[hexanes:ethyl acetate), 10:1 (v/v)]} in 11% yield. ##STR249##

To a stirred solution of 2.6 g (4.26 mmol) of Compound A13 in 80.0 mL ofdichloromethane was added 1.0 mL concentrated hydrochloric acid. Thesolution was stirred at room temperature for one hour, diluted with400.0 mL ethyl acetate, the organic solution washed four times with100.0 mL portions of a saturated aqueous solution of sodium chloridedried over 60.0 g of sodium sulfate, filtered, and concentrated underreduced pressure, at room temperature. The residue was purified on 300.0g of silica gel eluting with 2.0 L of 5:1 (v/v) hexanes:ethyl acetate.Evaporation of the solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 1.69 g (>4.26 mmol) of Compound A14 {R_(f) : 0.4[hexanes:ethyl acetate, 4/1 (v/v)]} that also contained a small amountof 4-methoxytriphenylmethyl chloride. The product was used in subsequentsynthetic reactions without further purification. ##STR250##

To a stirred solution of 0.55 g (1.62 mmol) of Compound A14 in 50.0 mLof anhydrous dichloromethane, under a nitrogen atmosphere, was added 3.0g of flame dried 3 Å molecular sieves (Aldrich Chemical Co.). Thesolution was stirred at room temperature for 15 minutes, 3.05 g (1.62mmol) of pyridinium dichromate (Aldrich Chemical Co.) was added in oneportion, the reaction mixture was stirred for an additional 40 minutes,the suspension was diluted with 50.0 mL dichloromethane, and the organicsuspension was washed first with 50 mL of a 10% (w/v) aqueous solutionof sodium thiosulfate and then with 50 mL of a saturted aqueous solutionof sodium chloride, dried over 60.0 g of sodium sulfate, filtered over20.0 g of Celite 545, and concentrated under reduced pressure at roomtemperature. The residue was purified on 50.0 g of silica gel elutingwith 300 mL of 7:1 (v/v) hexanes:ethyl acetate. Evaporation of thesolvent from the product-containing fractions (as identified by thinlayer chromatographic analysis) under reduced pressure at roomtemperature and drying for 30 minutes under vacuum at room temperatureprovided 0.46 g (1.37 mmol) of compound A15 {R_(f) : 0.88 [hexanes:ethylacetate, 2/1 (v/v)]} in 84% yield that was used directly in the nextreaction. ##STR251##

To a stirred solution of 0.46 g (1.37 mmol) of Compound A15 in 12.0 mLof tert-butyl alcohol and 3.0 mL (28.3 mmol) of 2-Methyl-2-butene(Aldrich Chemical Co.) at 0° C. was added dropwise 10.0 mL of an aqueoussolution containing 1.04 g (8.22 mmol) of sodium chlorite dihydrate(Eastman Kodak. Co., Rochester, N.Y.) and 1.11 g (8.04 mmol) of sodiumphosphate monobasic (Fisher Scientific Co.). The suspension was stirredat 0° C. for 20 minutes, the reaction mixture was quenched with 30.0 mLof a 10% (w/v) aqueous solution of sodium thiosulfate, diluted with100.0 mL of diethyl ether, and the resulting layers separated. Theorganic layer was washed with 50 mL of a saturated aqueous solution ofsodium chloride, dried over 60.0 g of sodium sulfate, filtered, andconcentrated under reduced pressure, at room temperature. The residuewas purified on 100.0 g of silica gel eluting first with 300 mL of 4:1(v/v) hexanes:ethyl acetate, then with 300 mL of 2:1 (v/v) hexanes:ethylacetate, and finally with 500 mL of 1:2 (v/v) hexanes:ethyl acetate.Evaporation of the solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) under reducedpressure, at room temperature, and drying overnight, under vacuum, atroom temperature provided 342.0 mg (0.97 mmol) of Compound A16 {R_(f) :0.28 [hexanes:ethyl acetate, 2:1 (v/v)]} {R_(f) : 0.45 [hexanes:ethylacetate, 2:1 (v/v)]} in 70.8% yield. ##STR252##

To a stirred solution of 342.0 mg (0.97 mmol) of Compound A16 in 16.0 mLof methyl alcohol and 0.5 mL (4.23 mmol) of quinoline (Aldrich ChemicalCo.) at room temperature was added 100.0 mg of 5% (wt/wt) palladium oncalcium carbonate, poisoned with lead (Aldrich Chemical Co.) under anitrogen atmosphere. The reaction mixture was evacuated under reducedpressure at room temperature, purged with hydrogen gas three times, andstirred under an atmosphere of hydrogen gas at atmospheric pressure fortwo and a half hours. The resulting reaction mixture was purged withnitrogen and filtered over 50.0 g of Celite 545 eluting with three 10.0mL portions of methyl alcohol. The filtrate was concentrated underreduced pressure at room temperature and diluted with 100.0 mL ofdichloromethane. The organic solution was washed twice with 60.0 mLportions of a 1.0N aqueous solution of hydrochloric acid and then oncewith 50.0 mL of a saturated aqueous solution of sodium chloride, driedover 60.0 g of sodium sulfate, filtered, and concentrated under reducedpressure at room temperature and dried over night under vacuum at roomtemperature to provide 340.0 mg (0.96 mmol) of crude Compound A17 {R_(f): 0.50 hexanes:ethyl acetate, 2/1 (v/v)]} in 99% yield. Compound A17 wasused in the next reaction without further purification. ##STR253##

To a stirred solution of 10.0 g (64.8 mmol) of decyn-1-ol (FarchanChemical Co., Gainesville, Fla.) in 10 mL of anhydrous pyridine, at 0°C., under a nitrogen atmosphere, was slowly added 18.5 g (97.0 mmol) ofpara-toluenesulfonyl chloride (99+%, Aldrich Chemical Co.) over afive-minute period. The reaction was allowed to warm slowly to roomtemperature, stirred for four hours, and diluted with 200.0 mL of ethylacetate. The organic solution was washed with 50.0 mL of a saturatedaqueous solution of sodium chloride, dried over 50.0 g of sodiumsulfate, filtered, concentrated under reduced pressure at roomtemperature and azeotroped to dryness with three 50.0 mL portions oftoluene using vacuum evaporation to provide 23.0 g of crude Compound A18{R_(f) : 0.06 [hexanes:ethyl acetate, 4:1 (v/v)]} which was used in thenext reaction without further purification. ##STR254##

To a stirred solution of 18.0 g (˜50.7 mmol) of Compound A18 in 240.0 mLof anhydrous dimethylsulfoxide (Fisher Scientific Co.) was added 36.0 g(194.6 mmol) of potassium phthalimide (Aldrich Chemical Co.) at roomtemperature under a nitrogen atmosphere. The reaction mixture was warmedto 50° C., stirred for three hours, and diluted with 1.0 L of ethylacetate. The resulting organic solution was washed first with 200.0 mLof a saturated aqueous solution of sodium bicarbonate and then with200.0 mL of a saturated aqueous solution of sodium chloride, dried over150.0 g of sodium sulfate, filtered, and concentrated under reducedpressure, at room temperature. The residue was purified on 300.0 g ofsilica gel eluting with 3.0 L of 6:1 (v/v) hexanes:ethyl acetate.Evaporation of the solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 13.0 g (45.8 mmol) of Compound A19 {R_(f) : 0.39[hexanes:ethyl acetate, 6/1 (v/v)]} in 84% yield. ##STR255##

To a stirred solution of 13.0 g (45.8 mmol) of Compound A19 in 200.0 mLof methyl alcohol and 8.1 mL (68.5 mmol) of quinoline at roomtemperature was added 1.0 g of 5% (wt/wt) palladium on calciumcarbonate, poisoned with lead under a nitrogen atmosphere. The reactionmixture was evacuated under reduced pressure, purged with hydrogen gasat room temperature three times, and stirred under an atmosphere ofhydrogen gas at atmospheric pressure for one hour. The resultingreaction mixture was purged with nitrogen and filtered over 100.0 g ofCelite 545 eluting with three 50.0 mL portions of methyl alcohol. Thefiltrate was concentrated under reduced pressure at room temperature anddiluted with 500.0 mL of dichloromethane. The organic solution waswashed with two 100.0 mL potions of a 1.0N aqueous solution ofhydrochloric acid and then with 100.0 mL of a saturated aqueous solutionof sodium chloride, dried over 150.0 g of sodium sulfate, filtered,concentrated under reduced pressure at room temperature and driedovernight under vacuum at room temperature to provided 13.0 g (45.6mmol) of crude Compound A20 {R_(f) :0.39 [hexanes:ethyl acetate, 6/1(v/v)]} in 99.6% yield. Compound A20 was used in the next reactionwithout further purification. ##STR256##

To a stirred solution of 6.0 g (21.0 mmol) of Compound A20 in 200.0 L ofabsolute ethyl alcohol (Quantum Chemical Co., Cincinnati, Ohio), at roomtemperature was added in 5.1 mL (105.0 mmol) of hydrazine hydrate (98%,Lancaster Chemical Co.). The reaction mixture was warmed to 75° C.,stirred for 75 minutes, cooled to room temperature, and diluted with300.0 mL of dichloromethane and 100 mL of water. The resulting layerswere separated, and the aqueous layer was extracted with two 50.0 mLportions of dichloromethane. The combined organic layers were dried over50.0 g of sodium sulfate, filtered, and concentrated under reducedpressure, at room temperature to provide 3.2 g of crude Compound A21{R_(f) : 0.08 [chloroform:methyl alcohol, 10:1 (v/v)]} in 100% yield.The crude product was used in the next reaction without furtherpurification. ##STR257##

To a mechanically stirred solution of 4.8 g (15.6 mmol) of Compound A4in 60.0 mL of anhydrous toluene, under a nitrogen atmosphere, at 0° C.,was added 6.0 mL (74.2 mmol) of anhydrous pyridine followed by thedropwise addition of 8.9 mL (17.2 mmol) of a 1.93M solution of phosgenein toluene over a 20-minute period. The mixture was stirred at 0° C. foran additional 15 minutes after which time 2.7 g (17.2 mmol) of CompoundA21 in 30.0 mL of anhydrous toluene was added dropwise over afive-minute period. The reaction mixture was stirred for an additional15 minutes, quenched with 30.0 mL of a saturated aqueous solution ofsodium bicarbonate, and diluted with 100.0 mL ethyl acetate. The organicsuspension was washed with 50.0 mL of a saturated aqueous solution ofsodium chloride, dried over 50.0 g of sodium sulfate, filtered, andconcentrated under reduced pressure at room temperature. The residue waspurified on 200.0 g of silica gel eluting with 3.0 L of 6:1 (v/v)hexanes:ethyl acetate. Evaporation of the solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 5.2 g (10.6mmol) of Compound A22 {R_(f) : 0.55 [hexanes:ethyl acetate), 10:1(v/v)]} in 68.3% yield. ##STR258##

To a mechanically stirred solution of 5.2 g (10.7 mmol) of Compound A22in 200 mL of glacial acetic acid in a three-necked Morton reaction flaskwas added 14.0 g (214.1 mmol) of activated zinc powder under a nitrogenatmosphere at room temperature. The reaction was stirred for 30 minutes,the suspension filtered through a pad of 60.0 g of Celite 545, andeluted with four 50.0 mL portions of methyl alcohol. The filtrate wasconcentrated under reduced pressure, at room temperature, and azeotropedto dryness with three 50 mL portions of toluene under vacuumevaporation. The crude yellow oil was purified on 200.0 g of silica gelby eluting with 2.0 L of 6:1 (v/v) hexanes:ethyl acetate and then with5.0 L of 1:1 (v/v) hexanes:ethyl acetate. Evaporation of the solventfrom the product-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure, at room temperature,and drying overnight, under vacuum, at room temperature provided 3.55 g(9.63 mmol) of Compound A23 {R_(f) : 0.08 [hexanes:ethyl acetate, 2:1(v/v)]} in 90% yield. ##STR259##

To a stirred solution of 23.7 g (0.118 mol) dodecanoic acid (AldrichChemical Co.) and 32.9 g (0.107 mol) of Compound A4 dissolved in 250.0mL anhydrous dichloromethane, at 0° C., was added 0.03 g (0.2 mmol)4-dimethylaminopyridine, then 29.2 g (0.143 mol)1,3-dicyclohexylcarbodimide. After being stirred for two and a halfhours at 25° C. the reaction mixture was diluted with 100.0 mL hexanes(200.0 mL), filtered and concentrated under reduced pressure at roomtemperature. The residue obtained was purified on a silica gel (2.0 Kg)column by elution with a 1:9 (v/v) mixture of ethyl acetate:hexanes.Evaporation of solvent from the product containing fractions (identifiedby use of thin layer chromatographic analysis) under reduced pressure atroom temperature and drying overnight under vacuum at room temperatureyielded 48.7 g (0.10 mol) of Compound A24 {R_(f) : 0.6 [hexanes:ethylacetate, 4:1 (v/v)]} in 84% yield. ##STR260##

To a mechanically stirred solution of Compound A24 (16.07 g, 32.99 mmol)dissolved in 150.0 mL of glacial acetic acid in a Morton flask at 0° C.was added 24.2 g (371.0 mmol) of zinc dust. After being warmed to 25° C.and stirred one hour, the reaction mixture was filtered through 50.0 gof Celite 545 and concentrated under reduced pressure at roomtemperature. The residue obtained was purified on silica gel (1.0 Kg)eluting first with ethyl acetate:hexanes [4:1 (v/v)], and then methylalcohol:chloroform:acetic acid [10:40:1 (v/v/v)]. Evaporation of solventfrom the product containing fractions (identified by use of thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature gave (10.8 g, 39.35mmol) of Compound A25 {R_(f) : 0.34 [hexanes:ethyl acetate, 2:1 (v/v)]}in 96% yield. ##STR261##

To a stirred solution of 1.0 mL (3.39 mmol) of 1-tridecyne (LancasterSynthesis) in 20.0 mL of anhydrous tetrahydrofuran at 0° C. under anitrogen atmosphere was added dropwise 1.5 mL (3.73 mmol) of 2.51Msolution of n-butyl lithium in hexanes over a five-minute period. Thereaction was stirred at 0° C. for one hour after which time the combinedsolution was transferred via a canula into a stirred solution of 0.46 mL(6.74 mmol) of methyl chloroformate (Aldrich Chemical Co.) in 10.0 mL ofanhydrous tetrahydrofuran at room temperature under a nitrogenatmosphere. The resulting reaction mixture was stirred for an additional30 minutes after which time the mixture was quenched with 10.0 mL of asaturated solution of ammonium chloride. The resulting mixture wasextracted with three 20 mL portions of ethyl acetate, and the combinedorganic layers were washed with one 10 mL portion of saturated sodiumchloride, dried over 20.0 g of anhydrous sodium sulfate, filtered andconcentrated under reduced pressure at room temperature. The crudeproduct, 0.69 g (2.89 mmol) of Compound A26 in 85.4%) yield, was used inthe next step without further purification after drying overnight undervacuum at room temperature {R_(f) : 0.78 [hexanes:ethyl acetate, 4:1(v/v)]}. ##STR262##

To a stirred suspension of 4.39 g (23.07 mmol) of copper(I) iodide(99.9%, Aldrich Chemical Co.) in 40.0 mL of anhydrous diethyl ether at0° C. under a nitrogen atmosphere was added dropwise 30.0 mL (46.2 mmol)of a 1.5M solution of methyl lithium in diethyl ether (Aldrich ChemicalCo.) over a 15-minute period until a clear, colorless solution wasobtained. The reaction solution was quickly transferred via a canulainto a stirred solution of 5.0 g (20.97 mmol) of Compound A26 in 50.0 mLof anhydrous diethyl ether at room temperature under a nitrogenatmosphere. After stirring for an additional five minutes, the resultingreaction mixture was quenched with 60.0 mL of a saturated solution ofammonium chloride and stirred for one hour. The resulting mixture wasdiluted with 50.0 mL of hexanes, filtered over a 50 g pad of Celite 545eluting with 50 mL of hexanes, and the layers separated. The aqueouslayer was extracted with two 50 mL portions of hexanes, and the combinedorganic layers were washed with one 50 mL portion of saturated sodiumchloride, dried over 100.0 g of anhydrous sodium sulfate, filtered andconcentrated under reduced pressure, at room temperature. The residuewas purified on two PrepPAK 500/silica cartridges (Waters Associates)connected in tandem, eluting with 10.0 L of a 98.5 to 2.5 (v/v) diethylether:hexanes solution at a 200 mL/minute flow rate using thePrepLC/System 500liquid chromatography device (Waters Associates) as thepumping and detection system. Evaporation of solvent from the productcontaining fractions (identified by use of thin layer chromatographicanalysis) under reduced pressure at room temperature and dryingovernight under vacuum, at room temperature yielded 1.25 g (4.93 mmol)of Compound A27 {R_(f) : 0.28 [diethyl ether:hexanes, 1:19 (v/v)]} in23.5% yield and 2.55 g (10.08 mmol) of Compound A28 {R_(f) : 0.22[diethyl ether:hexanes, 1:19 (v/v)]} in 48.1% yield. ##STR263##

To a stirred solution of 2.30 g (9.08 mmol) of Compound A27 in 8.0 mL ofanhydrous dichloromethane at 0° C. under a nitrogen atmosphere was addeddropwise 18.1 mL (18.1 mmol) of a 1.0M solution of diisobutylaluminumhydride in hexanes (Aldrich Chemical Co.) over a 15-minute period. Thereaction mixture was quenched with 60.0 mL of a saturated solution ofammonium chloride and stirred for an additional 45 minutes. Theresulting mixture was extracted with three 50 mL portions of ethylacetate, and the combined organic layers were washed with one 50 mLportion of a saturated solution of sodium chloride, dried over 100.0 gof anhydrous sodium sulfate, filtered and concentrated under reducedpressure, at room temperature. The residue was purified on 150.0 g ofsilica gel, eluting with 2.0 L of a 7:3 (v/v) mixture of hexanes anddiethyl ether. Evaporation of solvent from the product containingfractions (identified by use of thin layer chromatographic analysis)under reduced pressure at room temperature and drying overnight undervacuum at room temperature yielded 1.29 g (5,70 mmol) of Compound A29{R_(f) :0.19 [diethyl ether:hexanes, 3:7 (v/v)]} in 62.8% yield.##STR264##

To a stirred solution of 1.20 g (5.30 mmol) of Compound A28 in 6.0 mL ofchloroform at room temperature under a nitrogen atmosphere was added 4.6g (53.00 mmol) of activated manganese dioxide (Aldrich Chemical Co.) inone portion. The reaction suspension was refluxed for 30 minutes afterwhich time the mixture was cooled to room temperature, filtered over a50 g pad of Celite 545, eluting with 20 mL of chloroform. The combinedfiltrates were concentrated under reduced pressure, at room temperature.The resulting crude intermediate was dissolved in 50 mL oftetrahydrofuran and cooled to 0° C., after which time 4.0 mL of2-methyl-2-butene was added in one portion. The reaction solution wastreated with a dropwise addition of 10.0 mL of a 1:0.09:1 ratio (w/w/v)of sodium phosphate dibasic (Fisher Scientific Co.), sodium chlorite(Eastman Kodak Co.), and water over a five-minute period. The reactionmixture was stirred for an additional 30 minutes at 0° C. after whichtime the mixture was quenched with 50.0 mL of a 10% solution of sodiumthiosulfate and stirred for an additional 10 minutes. The resultingmixture was acidified to a pH of 3.0 using 1.0N aqueous hydrochloricacid and extracted with three, 50 mL portions of ethyl acetate. Thecombined organic layers were washed with one, 50 mL portion of asaturated solution of sodium chloride, dried over 100.0 g of anhydroussodium sulfate, filtered and concentrated under reduced pressure, atroom temperature. The residue was purified on 200.0 g of silica gel,eluting with 2.0 L of a 3:1 (v/v) mixture of hexanes and diethyl ether.Evaporation of solvent from the product-containing fractions (identifiedby use of thin layer chromatographic analysis) under reduced pressure,at room temperature and drying overnight under vacuum at roomtemperature yielded 1.01 g (4.22 mmol) of Compound A30 {R_(f) :0.21[diethyl ether:hexanes, 3:7 (v/v)]} in 79.6% yield. ##STR265##

To a stirred solution of 20.8 mg (0.09 mmol) of Compound A30 in 5.0 mLof anhydrous dichloromethane at 0° C. under a nitrogen atmosphere wasadded 15.7 μL (0.18 mmol) of oxalyl chloride (Aldrich Chemical Co.)dropwise over a two minute period. The reaction mixture was stirred for40 minutes at 0° C. after which time the mixture was concentrated underreduced pressure at room temperature under anhydrous conditions anddried for one hour under vacuum at room temperature to provide CompoundA31 as a crude syrup which was used in the next reaction without furtherpurification. ##STR266##

To a stirred solution of 1-octyne (31.6 g, 0.287 mol; Aldrich ChemicalCo.) in anhydrous tetrahydrofuran (250.0 mL), n-butyllithium (163.5 mL,0.315 mol) was added dropwise a 0° C. under a nitrogen atmosphere over a40-minute period. The solution was stirred at 25° C. for one hour,1,3-diiodopropane (103.0 g, 0.349 mol; Aldrich Chemical Co.) was addeddropwise over a 10-minute period, and the resulting mixture was stirredfor 20 hours. The completed reaction mixture was diluted with 250.0 mLhexanes and poured into 400.0 mL of ice water. The product was washedtwice with 300.0 mL portions of saturated aqueous sodium chloridesolution, dried over 500.0 g sodium sulfate, filtered, and concentratedunder reduced pressure at room temperature. The residue was purified ona silica gel (1.0 kg) column, eluting with hexanes. Evaporation ofsolvent from the product-containing fractions (as determined by thinlayer chromatographic analysis) under reduced pressure at roomtemperature provided 59.95 g (0.21 mol) of Compound B1 {R_(f) :0.8[hexanes]} in 78% yield. ##STR267##

To a stirred solution of potassium cyanide (55.0 g, 0.845 mol; AldrichChemical Co.) in dimethylsulfoxide (750.0 mL) was added dropwise 135.0 g(0.485 mol) of Compound B1 over a 30-minute period. The solution wasthen stirred five hours at 50° C., diluted with 250.0 mL hexanes, andwashed with 250.0 mL water. The organic layer was dried over 50.0 gmagnesium sulfate, filtered, and concentrated at room temperature underreduced pressure. The residue was purified on a silica gel (2.0 kg)column, eluting with 95:5(v/v) hexanes/ethyl acetate. Evaporation ofsolvent from the product-containing fractions (as determined by thinlayer chromatographic analysis) under reduced pressure at roomtemperature provided 51.4 g (0.29 mol) of Compound B2 {R_(f) :0.3 [ethylacetate:hexanes, 5:95(v/v)]} in 81% yield. ##STR268##

Compound B2 (9.36 g, 0.053 mol) was dissolved in ethylene glycol (90.0mL; Aldrich Chemical Co.) and, to the solution, was added 8.89 g (0.158mol) potassium hydroxide (Fisher Scientific). After stirring for fourhours at 140° C. and then cooling to 25° C., the reaction mixture wasdiluted with 90.0 mL water and then washed twice with 90.0 mL portionsof dichloromethane. The aqueous layer was acidified with 200.0 mL 1Nhydrochloric acid, and the produce was extracted with hexanes (250.0mL). The extract was dried over 50 g magnesium sulfate, filtered, andconcentrated under reduced pressure at room temperature to provide 8.58g (0.04 mol) of Compound B3 {R_(f) :0.2 [hexanes:ethyl acetate,4:1(v/v)]} in 82% yield. ##STR269##

Compound B3 (20.0 g, 0.102 mol) and Lindlar catalyst (i.e., 5% palladiumon calcium carbonate, poisoned with lead; 86.0 g) were added to asolution of quinoline (10.0 mL, 0.084 mol) in hexanes (190.0 mL). Thereaction mixture was stirred under hydrogen gas for five hours,filtered, and evaporated. The residue was diluted with 10.0 mLdichloromethane, made basic with 150.0 mL of 1N sodium hydroxide, andthe aqueous layer was washed with 50.0 mL of dichloromethane. Theaqueous layer was then acidified with 20.0 mL 6N hydrochloric acid, andextracted with ethyl acetate (200.0 mL). The extract was washed with200.0 mL saturated aqueous sodium chloride solution, dried over 100.0 gsodium sulfate, filtered, and concentrated under reduced pressure toprovide 19.8 (0.1 mol) of Compound B4 {R_(f) :0.2 [hexanes:ethylacetate, 4:1(v/v)]} in 98% yield. ##STR270##

Compound B4 (23.7 g, 0.119 mol) and Compound A4 (32.9 g, 0.107 mol) weredissolved in 250.0 mL anhydrous dichloromethane at 0° C. and to thesolution was first added 0.03 g (0.2 mmol) 4-dimethylaminopyridinefollowed by 29.2 g (0.143 mol) 1,3-dicyclohexylcarbodimide. The solutionwas stirred for two and a half hours at 25° C., diluted with hexanes(100.0 mL), filtered, and concentrated under reduced pressure at roomtemperature. The residue was purified on a silica gel (2.0 kg) column,eluting with 1:9(v/v) ethyl acetate/hexanes. Evaporation of solvent fromthe product-containing fractions (as determined by thin layerchromatographic analysis) under reduced pressure at room temperatureprovided 48.7 g (0.10 mol) of Compound B5 {R_(f) :0.6 [hexanes:ethylacetate, 4:1(v/v)]} in 84% yield. ##STR271##

Compound B5 (16.1 g, 0.412 mol) was dissolved in 150.0 mL glacial aceticacid and to the solution was added 24.2 g (0.370 mol) since metal powderat 0° C. The reaction mixture was stirred vigorously for 40 minutes at25° C., diluted with 150.0 mL ethyl acetate, filtered, and concentratedunder reduced pressure at room temperature. The residue was purified ona silica gel (1.0 kg) column and eluted with 9:1(v/v) hexanes/ethylacetate. Evaporation of solvent from the product-containing fractions(as determined by thin layer chromatographic analysis) under reducedpressure at room temperature provided 10.8 g (0.04 mol) of Compound B5{R_(f) :0.3 [hexanes:ethyl acetate, 4:1(v/v)]} in 96% yield. ##STR272##

To 101.0 g (1.54 mol) of activated zinc in 500.0 mL refluxing anhydroustetrahydrofuran was added 3.0 mL of ethyl bromoacetate and 67.5 mL(0.308 mol) of undecyl cyanide (in one portion; Aldrich Chemical Co.).To the resulting mixture was added dropwise 134.0 mL (1.232 mol) ofethyl bromoacetate over three hours. The mixture was refluxed for anadditional 10 minutes, cooled to room temperature, and the reactionquenched by the slow addition of 160.0 mL of saturated aqueous potassiumcarbonate solution. The resultant heterogeneous mixture was rapidlystirred for 30 minutes and then filtered through 500.0 g Celite 545,yielding a clear, yellow solution of crude enamino ester. The solutionwas then acidified with 300.0 mL of 1.0N hydrochloric acid, stirred forthree hours, diluted with 1.0 L hexanes, and neutralized by addition of1.0 L saturated aqueous sodium hydrogen carbonate. The organic layer wasthen washed with 400.0 mL saturated aqueous sodium chloride solution,dried over 1.0 kg sodium sulfate, filtered, and concentrated underreduced pressure. The residue was purified on silica gel (1.0 kg),eluting with 6:1(v/v) hexanes/ethyl acetate to provide 80.6 g (0.298mol) of Product C1 {R_(f) :0.7 [hexanes:ethyl acetate, 4:1(v/v)]} in 97%yield. ##STR273##

[R]-2,2'-Bis(diphenylphosphino)-1,1'-binaphthyl ruthenium dichloridehemi triethylamine complex, as a clear orange-brown solution, wasprepared form [R]-(+)-2,2'-Bis(diphenylphosphino)-1,1'-binaphythyl andcyclooctadienyl ruthenium dichloride as described above.

A solution of 311.0 g (1.15 mol) of Compound C1 in 330.0 mL anhydrousmethyl alcohol was deoxygenated by three freeze-thaw vacuum degassingcycles in liquid nitrogen under a nitrogen atmosphere. The solution of[R]-2,2'-Bis(diphenylphosphino)-1,1'-binaphythyl ruthenium dichloridehemi triethylamine complex catalyst was added to the CompoundC1-containing solution using a syringe. Using a catheter under argon,the reaction mixture was pumped into an argon-flushed 2.0 Lhydrogenation bomb containing 3.0 g methyl alcohol-washed Dowex 50×8-200H⁺ resin. The bomb was charged to 1480 psi with hydrogen gas, and thereaction mixture was stirred, at 25° C., for 66 hours. When the pressurehad dropped 300 psi, excess hydrogen gas was vented, the reactionmixture was filtered, and the volatiles were removed under reducedpressure to provide 310.2 g (1.14 mol) of Compound C2 in 99% yield.##STR274##

Compound C2 (94.4 g, 0.347 mol) was dissolved in tetrahydrofuran (800.0mL) and, to the solution, was added 2.5M aqueous sodium hydroxide (300.0mL, 0.75 mol). The resultant mixture was stirred vigorously under anitrogen atmosphere, at 25° C., for one and a half hours. The reactionmixture was diluted with 1.0 L of 1:1(v/v) diethyl ether/hexanes and theaqueous layer separated. The organic phase was further extracted with200.0 mL of water, the aqueous phases combined, and the combined aqueousphases acidified with 20.0 mL of 6N hydrochloric acid. The acidifiedmixture was then extracted with 2.0 L diethyl ether and the extractwashed first with 1.0 L water, and then with 500.0 mL saturated aqueoussodium chloride solution, and dried over 200.0 g magnesium sulfate.Solvent was concentrated under reduced pressure at room temperature, andthe grayish solid obtained was dissolved in 2.0 L hot 60° C.acetonitrile. To the solution at 60° C. was added dicyclohexylamine(80.0 mL, 0.40 mol), and the resultant mixture cooled to -20° C., toprovide 102.1 g (0.24 mol) of Compound C3 {R_(f) :0.38 [hexanes:ethylacetate:glacial acetic acid, 1:1:0.1 (v/v/v)]}, as slightly off-while,fine needles in 71% yield. ##STR275##

Compound C3 (102.1 g, 0.24 mol) was suspended in 2.0 L ethyl acetate,under a nitrogen atmosphere, at 0° C. To the suspension was added 37.2 g(369.0 mmol) triethylamine, followed by 48.9 g (246.0 mmol)2-bromoacetophenone (in one portion). After three hours, the reactionmixture was warmed to room temperature and stirred for an additional sixhours. The reaction mixture was then vacuum-filtered. The residue waswashed with 400.0 mL ethyl acetate. The filtrate was washed first with500 mL 0.8M hydrochloric acid, then with 500.0 mL water, and finallywith 200.0 mL saturated aqueous sodium chloride solution, and dried over300.0 g magnesium sulfate. The solvent was evaporated at 50° C., underreduced pressure, to yield a gray congealed solid which wasrecrystallized from 1.1 L hexanes and dried in a vacuum oven at 50° C.to provide 81.09 g (223.0 mmol) of Compound C4 {R_(f) :0.65[chloroform:methyl alcohol, 95:5 (v/v)]} as an off-white solid in 91%yield. ##STR276##

To a suspension of 2.5 g (9.18 mmol) C4 and 1.0 g 4A molecular sieves in28.0 mL of 3:1(v/v) hexanes/dichloromethane, under a nitrogenatmosphere, was added 3.8 mL (13.4 mmol) of4-methyoxybenzyltrichloroimidate (prepared by the method of Audia etal., J Org Chem. 1989 54:3738). The reaction mixture was cooled to 0°C., and 63.0 μL (0.51 mol) neat boron trifluoride etherate was addeddropwise. After five minutes the reaction was quenched with 2.0 mLsaturated aqueous sodium bicarbonate solution and the reaction mixturewarmed to 25° C. The mixture was then extracted with 100.0 mL ethylacetate, washed with 50.0 mL saturated aqueous sodium chloride solution,dried over 50.0 g sodium sulfate, filtered, and the solvent concentratedunder reduced pressure at room temperature. The residue was purified ona silica gel (300.0 g) column, eluting with 1:9(v/v) ethylacetate/hexanes. Evaporation of solvent from the product-containingfractions (as determined by thin layer chromatographic analysis) underreduced pressure at room temperature provided 3.1 g (7.97 mmol) ofCompound C5 {R_(f) :0.7 [ethyl acetate:hexanes, 3:17(v/v)]} in 87%yield. ##STR277##

Compound C5 (3.1 g) was dissolved in tetrahydrofuran (30.0 mL) and tothe solution was added 16.0 mL (40.0 mmol) 2.5N sodium hydroxide. Thereaction mixture was stirred for six days at 25° C., diluted with 100.0mL hexanes, and the pH adjusted to 5.0 with 40.0 mL 1N hydrochloricacid. The reaction mixture was then extracted with 300.0 mL ethylacetate, washed with 100.0 mL saturated aqueous sodium chloridesolution, dried over 200.0 g sodium sulfate, filtered, and the solventconcentrated under reduced pressure at room temperature. The residue waspurified on a silica gel (300.0 g) column, eluting with a gradient ofmethyl alcohol/chloroform [3:17(v/v) to 1:9(v/v)] Each gradient mixturealso contained two drops of glacial acetic acid per 100.0 mL solvent. Atotal of 1.46 g (4.0 mmol) of Compound C6 {R_(f) :0.14 [hexanes:ethylacetate, 2:1(v/v)]} was provided in 50% yield. ##STR278##

Compound C1 (2.04 g, 7.96 mmol) was dissolved in 25.0 mL dry diethylether. To this solution, 1,3-propanedithiol (0.8 mL, 7.97 mmol; in oneportion; Aldrich Chemical Co.) was added, under a nitrogen atmosphere,at 0° C. Next, 1.0 mL (8.13 mmol) of boron trifluoride etherate wasadded dropwise over a two minute period. The reaction mixture wasstirred at 0° C., for 30 minutes at room temperature for 48 hours,poured into a saturated aqueous sodium bicarbonate (200.0 mL) solution,and stirred for an additional 30 minutes. The solution was extractedthree times with 50.0 mL portions of hexanes. The combined organicfractions were washed with 50.0 mL of saturated aqueous sodium chloridesolution, dried over 50.0 g magnesium sulfate, filtered, andconcentrated under reduced pressure at room temperature. The product waspurified on a silica gel (200.0 g) column, eluting with 2.0 L of a 0 to5:95(v/v) gradient of ethyl acetate/hexanes. The fractions containingpurified Compound C7 {R_(f) :0.44 [hexanes:ethyl acetate, (95.5)]} wereconcentrated and used in subsequent synthetic reactions. ##STR279##

Compound C7 2.88 g (8.01 mmol) was dissolved in tetrahydrofuran (20.0mL) and, to the solution, 10.0 mL of 2.5M aqueous sodium hydroxidesolution was added. The reaction mixture was warmed to 100° C., stirredfor 16 hours, cooled to room temperature, adjusted to pH 2.0 with 10.0mL 1.0N hydrochloric acid, and extracted with 200.0 mL ethyl acetate.The organic layer was washed with 50.0 mL saturated aqueous sodiumchloride solution, dried over 50.0 g sodium sulfate, filtered, andconcentrated under reduced pressure at room temperature. Purificationwas accomplished on a silica gel (300.0 g) column, eluting first with1.0 L of 4:1(v/v) hexanes/ethyl acetate and then with 1.0 L of9:1:0.1(v/v/v) chloroform/methyl alcohol/glacial acetic acid.Evaporation of solvent from the product-containing fractions (asdetermined by thin layer chromatographic analysis) under reducedpressure at room temperature provided 2.10 g (6.33 mmol) of Compound C8{R_(f) :0.20 [hexanes:ethyl acetate, 2:1 (v/v)]} in 79% yield.##STR280##

To a mechanically stirred suspension of 1004.0 g (15.4 mol) of activatedzinc powder in 2.5 L of anhydrous tetrahydrofuran under a nitrogenatmosphere at room temperature was added dropwise 30.0 mL (0.19 mol) ofbenzyl bromoacetate over a 10-minute period. The reaction mixture waswarmed to reflux, 712.0 mL (3.25 mol) of n-undecyl cyanide (AldrichChemical Co.) added dropwise over a 15-minute period, and an additional1.00 L (6.3 mol) of benzyl bromoacetate (Aldrich Chemical Co.) addedover a four-hour period. After an additional hour of refluxing, thereaction mixture was cooled to room temperature and slowly poured into3.0 L of a stirred saturated aqueous solution of potassium carbonate. Tothe resulting solution was added 1.0 kg of Celite 545 and theheterogeneous mixture filtered through a pad of 200.0 g of Celite 545 byelution with four 1.0 L portions of ethyl acetate. The filtrate wasseparated, and the aqueous layer extracted with two 500.0 mL portions ofethyl acetate. The combined organic layers were washed with a 500.0 mLportion of a saturated aqueous solution of sodium chloride, dried oversodium sulfate 100 g, filtered, and concentrated under vacuum todryness. The crude orange oil was vigorously stirred at room temperaturein a two-phase system of 1.0 L of hexanes and 1.0 L of 1.0N hydrochloricacid with a dropwise addition of 80 mL concentrated hydrochloric acidover a three-hour period. After stirring the final heterogeneoussolution for an additional 20 minutes, the layers were separated, andthe aqueous layer was extracted with two 200.0 mL portions of hexanes.The combined organic layers were washed first with 500.0 mL of asaturated aqueous solution of sodium hydrogen carbonate and then with500.0 mL of a saturated aqueous solution sodium chloride, dried oversodium sulfate 100 g (filtered, and concentrated under vacuum todryness. The crude orange liquid was purified over 2.5 kg of silica geleluting with 12.0 L of 9:1(v/v) hexanes:ethyl acetate. Evaporation ofsolvent form the product-containing fractions (as identified by thinlayer chromatographic analysis) under reduced pressure at roomtemperature and drying overnight under vacuum at room temperatureprovided 884.7 g (2.66 mol) of Compound D1 {R_(f) :0.67 [hexanes:ethylacetate, 4:1(v/v)]} as a yellow solid in 82% yield. ##STR281##

To a stirred solution of 17.9 g (53.9 mmol) of Compound D1 in 110.0 mLof methyl alcohol, under a nitrogen atmosphere, was added 0.9 g of 20%palladium hydroxide on carbon (Aldrich Chemical Co.). The resultingsuspension was purged with hydrogen gas and evacuated under reducedpressure three times followed by stirring under an atmosphere ofhydrogen gas at atmospheric pressure and room temperature for one hour.The completed reaction mixture was diluted with 100 mL ofdichloromethane, filtered over a 50.0 g pad of Celite 545 and theresulting filter cake washed with two 50.0 mL portions ofdichloromethane. The combined filtrates were concentrated under reducedpressure, at room temperature. The crude product was quickly purifiedover 200.0 g of silica gel eluting with 2.0 L of 9:1(v/v)chloroform:methyl alcohol. Evaporation of the solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying for 30 minutes under vacuum at room temperature provided 11.5 g(47.5 mmol) of Compound D2 {R_(f) :0.56 [chloroform:methylalcohol:acetic acid, 9:1:0.1 (v/v/v)]} as a white solid in 88% yield.Compound D2 was used immediately in the next reaction to avoiddecomposition. ##STR282##

To a vigorously stirred solution of 1000.0 g (9.42 mol) ofmethylthioglycolate (Aldrich Chemical Co.) in 2.0 L of anhydroustetrhydrofuran, under a nitrogen atmosphere, at 0° C., was added 1312.0ml (9.41 mol) of triethylamine followed by the dropwise addition of2138.0 mL (9.42 mol) of 1-iodoundecane (Aldrich Chemical Co.) over a 12hour period. The reaction mixture was warmed to room temperature,stirred for an additional 24 hours, diluted with 2.0 L of ethyl acetate,and washed first with 1.0 L of a 1.0N aqueous solution of hydrochloricacid, then with 1.0 L of a saturated aqueous solution of sodiumbicarbonate, and finally with 1.0 L of a saturated aqueous solution ofsodium chloride. The organic layer was dried over 500.0 g of sodiumsulfate, filtered, and concentrated under reduced pressure, at roomtemperature to provide 2230.0 g of Compound E1 {R_(f) :0.69[hexanes:ethyl acetate, 4:1(v/v)]} which was used in the next stepwithout further purification. ##STR283##

To a stirred solution of 2100.0 g (˜8.07 mol) of crude Compound E1 in5.2 L of acetone and 5.2 L of water at -10° C. was added portion-wise5.0 kg (8.13 mol) of potassium peroxymonosulfate (OXONE, AldrichChemical Co.) over a three-hour period using a powder addition funnel.The mixture was warmed to 0° C., stirred for an additional two hours,and slowly quenched at 0° C. with 3.0 L of a 3.0M aqueous solution ofsodium thiosulfate. The mixture was diluted with 8.0 L ofdichloromethane and 4.0 L of water, filtered through a 500.0 g pad ofCelite 545, and the resulting filter cake eluted with three 500.0 mLportions of chloroform. The combined filtrates were washed first with10.0 L of a saturated aqueous solution of sodium bicarbonate and thenwith 20.0 L of a saturated aqueous solution of sodium chloride, and theorganic layer dried over 2.5 kg of sodium sulfate, filtered, andconcentrated under reduced pressure at room temperature. The crudeproduct was purified by washing the resulting solid with 8.0 L ofhexanes to provide 1541.0 g (5.58 mol) of Compound E2 [melting point(m.p.) 62.6°-63.5° C.] {R_(f) :0.2 [hexanes:ethyl acetate), 1:1(v/v)]}as a white solid in 69% yield. ##STR284##

To a stirred mixture of 1644.0 g (5.95 mol) of Compound E2 in 6.0 L oftoluene and 48.0 L of an aqueous solution of 0.05M phosphate buffer wasadded 12.0 g of lipase (PS-800, Amano Intl. Enzyme Co., Troy, Va.) atroom temperature. The reaction mixture was stirred at room temperaturefor 24 hours, an additional 11.9 g of lipase (PS-800) added, and thefinal suspension stirred for 96 hours. The resulting mixture wasacidified to pH 1.0 with approximately 4.0 L of an aqueous solution of1.0N hydrochloric acid, diluted with 6.0 L of chloroform, and the layersseparated. The aqueous layer was extracted with five 1.0 L portions ofchloroform and the combined organic layers were dried over 500.0 g ofsodium sulfate, filtered, and concentrated under reduced pressure atroom temperature. The crude product was suspended in 6.0 L of ethylacetate, warmed to 60° C., cooled to room temperature, filtered, and theresulting solid washed with two 500.0 mL portions of ethyl acetate. Thecrude solid was recrystalized twice from 10.0 L of ethyl acetate toprovide 305.0 g (1.16 mol, m.p. 80.4°-81.8° C.) of Compound E3 in 19.5%yield. The combined mother liquors were concentrated under reducedpressure at room temperature and purified over 2.0 kg of silica geleluting first with 4.0 L of hexanes, then with 6.0 L of 4:1(v/v)hexanes:ethyl acetate, then with 4.0 L of chloroform, and finally with10.0 L of 9:1:0.1(v/v/v) chloroform:methyl alcohol:acetic acid.Evaporation of the solvent from the product-containing fractions (asidentified by thin layer chromatographic analysis) under reducedpressure at room temperature and drying overnight under vacuum at roomtemperature provided 90.0 g (0.34 mol, 5.8% yield) of E3 {R_(f) :0.12[chloroform:methyl alcohol:acetic acid, 9:1:0.1 (v/v/v)]}, 300.0 g of amixture of E3 and E4, and 500.0 g (1.81 mol, 30.4% yield) of E4 {R_(f):0.63 [chloroform:methyl alcohol:acetic acid, 9:1:0.1 (v/v/v)]}.##STR285##

To a solution of 500.0 g (1.81 mol) of Compound E4 in 5.0 L of methylalcohol at room temperature was added dropwise 2.0 L of a 2.0N aqueoussolution of sodium hydroxide until a pH range of 11 to 12 was reached.The mixture was stirred for one hour, the mixture acidified to a pH of2.0-3.0 with 2.0 L of a 2.0N aqueous solution of hydrochloric acid, anddiluted with 4.0 L of chloroform. The layers were separated, and theaqueous layer was extracted with three 1.0 L portions of chloroform. Thecombined organic layers were dried over 500.0 g of sodium sulfate,filtered, and concentrated under reduced pressure, at room temperature.The crude solid was recrystalized four times from 6.0 L of ethyl acetateto provide 167.0 g (0.64 mol, m.p. 81.2°-82.1° C.) of pure E5 {R_(f):0.12 [chloroform:methyl alcohol:acetic acid, 9:1:0.1 (v/v/v)]} in 35.2%yield. Concentration of the mother liquors under reduced pressure atroom temperature provided an additional 250.0 g of crude E5. ##STR286##

To a stirred solution of 1.07 g (3.85 mmol) of crude Compound E1 in 20.1mL of anhydrous dichloromethane at 0° C. was added portionwise 2.86 g(7.7 mmol of 3-chloroperoxybenzoic acid over a 10-minute period. Themixture was stirred for one hour and slowly quenched at 0° C. with 10.0mL of a 3.0M aqueous solution of sodium thiosulfate. The mixture wasthen diluted with 100.0 mL of a saturated aqueous solution of sodiumbicarbonate, 100.0 mL of a saturated aqueous solution of sodiumchloride, and the resulting organic layer was dried over 100.0 g ofsodium sulfate, filtered, and concentrated under reduced pressure atroom temperature. The crude product was purified by crystallization fromhexanes to provide 970.0 mg (3.32 mmol) of Compound E6 {R_(f) :0.67[hexanes:ethyl acetate, 1:1 (v/v)]}. ##STR287##

To the solution of 970.0 mg (3.32 mmol) of Compound E6 in 15.0 mL ofmethyl alcohol at room temperature was added 4.0 mL of a 1.0N aqueoussolution of sodium hydroxide. The mixture was stirred for one hour afterwhich time the resulting mixture was acidified with 4.0 mL of a 2.0Naqueous solution of hydrochloric acid. The final mixture was dilutedwith 100.0 mL of chloroform, the layers were separated, and the aqueouslayer was extracted with three 100.0 mL portions of chloroform. Thecombined organic layers were dried over 100.0 g of sodium sulfate,filtered, and concentrated under reduced pressure at room temperature.The crude solid Compound E7 was used without further purification.##STR288##

To a refluxing, mechanically stirred suspension of 5.5 g (84.1 mmol) ofactivated zinc powder in 50 mL of anhydrous tetrahydrofuran was addeddropwise 1.0 mL (9.0 mmol) of ethyl bromoacetate over a one-minuteperiod, under a nitrogen atmosphere. 3.0 g (16.9 mmol) of Compound B2was then added in one portion followed by the dropwise addition of 7.4mL (66.7 mmol) of ethyl bromoacetate over a 45 minute period. After 10minutes of refluxing, the reaction mixture was cooled to roomtemperature, diluted with 170.0 mL of tetrahydrofuran, and quenched withthe dropwise addition of 22.0 mL of a 50% saturated aqueous solution ofpotassium carbonate over a 10-minute period. The resulting suspensionwas stirred for 30 minutes (after which time stirring was discontinued),the tetrahydrofuran solution decanted from the zinc solid, and the zincsolid washed with four additional 50.0 mL portions of tetrahydrofuran.The resulting product solutions were combined and vigorously stirredwith 17.0 mL of 1.0N hydrochloric acid, stirred for two hours, andconcentrated under reduced pressure, at room temperature. The residuewas dissolved in 200.0 mL of dichloromethane and the organic solutionwashed with a 50.0 mL portion of a saturated aqueous solution of sodiumbicarbonate, dried over 50.0 g of sodium sulfate, filtered, andconcentrated under reduced pressure at room temperature. The crudeproduct was purified over 200.0 g of silica gel eluting with 3.0 L of10:1(v/v) hexanes:ethyl acetate. Evaporation of solvent from theproduct-containing fractions (as identified by thin layerchromatographic analysis) under reduced pressure at room temperature anddrying overnight under vacuum at room temperature provided 3.52 g (13.2mmol) of Compound G1 {R_(f) :0.65 [hexanes:ethyl acetate, 4:1(v/v)]} asa clear, colorless oil in 78.2% yield. ##STR289##

To a stirred solution of 1.5 g (5.64 mmol) of Compound G1 in 7.0 mL ofanhydrous diethyl either and 434.0 μL (6.22 mmol) mercaptoethanol(Aldrich Chemical Co.), at 0° C., under a nitrogen atmosphere, was addeddropwise 762.0 μL (6.20 mmol) boron trifluoroetherate over a five-minuteperiod. The mixture was warmed to room temperature, stirred at roomtemperature for 16 hours, cooled to 0° C., quenched with 10.0 mL of asaturated aqueous solution of sodium bicarbonate, and stirred for anadditional five minutes. The resulting mixture was extracted with three50.0 mL portions of diethyl ether, and the combined organic layers werewashed with 50.0 mL saturated aqueous sodium chloride, dried over 50.0 gof sodium sulfate, filtered, and concentrated under reduced pressure atroom temperature. The crude product was dissolved in 14.0 mL oftetrahydrofuran, mixed with 7.0 mL of a 2.5M aqueous solution of sodiumhydroxide, and stirred at 80° C. for 16 hours. The final reactionmixture was cooled to room temperature, extracted with three 10.0 mLportions of diethyl ether, and the aqueous layer acidified to a pH of2.0 with a 1.0N aqueous solution of hydrochloric acid. The resultingaqueous suspension was extracted three 10.0 mL portions of diethylether, and the combined organic layers were washed with 10.0 mLsaturated aqueous sodium chloride, dried over 20.0 g of sodium sulfate,filtered, concentrated under reduced pressure at room temperature anddried overnight under vacuum at room temperature to provide 1.40 g (4.68mmol) of crude Compound G2 {R_(f) :0.6 [hexanes]} in 83% yield. Theproduct was used in the next reaction without further purification.

There now follow a characterization of the compounds described hereinand a description of assays which are used to test their efficacy. Theseexamples are provided to illustrate, not limit, the invention.

EXAMPLE 2 Compound Characterization Compound 2

¹ H NMR (CDCl₃) δ:6.08 ppm (1 H.sub.α,d,J=1.71 Hz), 5.8(1H.sub.β,d,J=1.22 Hz), 5.48-5.10(3 H,m), 4.32-3.80(3 H,m), 2.33-1.92(15H,m.s).

Compound 3

¹ H NMR (CDCl₃) δ:7.52-7.30 ppm (5 H,m), 5.52(2 H,d), 5.32(2 H,m),4.61(1 H,m), 4.33(1 H,dd), 4.12(1 H,dd), 2.18(3 H,s), 2.09(3 H,s),2.05(3 H,s), 2.03(3 H,s).

Compound 4

¹ H NMR (CDCl₃) δ:7.48-7.12 ppm (5 H,m), 5.51(1 H,s), 4.20(1 H,s),4.02(1 H,s), 3.97-3.68 (4 H,m).

Compound 5

¹ H NMR (CDCl₃) δ:7.49-7.39 ppm (5 H,m), 5.78(1 H,s), 4.39(1 H,d),4.26(1 H,dd), 4.01(1 H,ddd), 3.81(1 H,dd), 3.77(2 H,m), 1.52(3 H,s),1.50(3 H,s), 1.43(3 H,s), 1.37(3 H,s).

Compound 6

¹ H NMR (CDCl₃) δ:6.30 ppm (1 H,dd, J=1.50,6.11 Hz), 4.72(1H,dd,J=1.79,6.07 Hz), 4.34(1 H,d,J=7.39 Hz), 3.94(1 H,dd,J=5.48,10.92Hz), 3.84-3.71(3 H,m), 1.53(3 H,s), 1.43(3 H,s).

Compound 7

¹ H NMR (CDCl₃) δ:6.33 ppm (1 H,dd,J=1.46,6.1 Hz), 5.32(1H,dt,J=1.7,1.7,7.8 Hz), 4.76(1 H,dd,J=2.0,6.1 Hz), 4.03(1H,dd,J=1.71,7.81 Hz), 3.97(1 H,m), 3.87-3.76(2 H,m), 2.09(3 H,s), 1.52(3H,s), 1.41(3 H,s).

Compound 8

R_(f) :0.21 [methyl alcohol:chloroform, 5:95 (v/v)] ¹ H NMR (CDCl₃)δ:5.61 ppm (1 H,d,J=8.79 Hz), 5.13(1 H,t,J=9.5 Hz), 3.91(1H,dd,J=5.37,10.98 Hz), 3.75(1 H,t,J=10.25 Hz), 3.68(1 H,t,J=9.76 Hz),3.56(1 H,dd,J=8.79.9.52 Hz), 3.49(1 H,m), 2.15(3 H,s), 1.46(3 H,s),1.38(3 H,s).

Compound 9

¹ H NMR (CDCl₃) δ:5.46 ppm (1 H.sub.β,t,J=10.0 Hz), 5.34(1 H.sub.α,t),5.00(1 H.sub.β,t,J=9.7 Hz), 4.78(1 H.sub.α,dd,J=5.1,7.8 Hz),4.04-3.64(mH), 3.41-3.23(mH),2.12(3 H,s), 1.45(3 H.sub.α,s), 1.44(3H.sub.α,s), 1.39(1 H.sub.β,s), 1.35(3 H.sub.β,s).

Compound 10a

¹ H NMR (CDCl₃) δ:8.78 ppm (1 H,s), 6.42(1 H,d,J=3.0 Hz), 5.48(1H,t,J=9.52 Hz), 3.95-3.86(2 H,m), 3.82-3.79(2 H,m), 3.59(1H,dd,J=3.67,10.3 Hz), 2.15(3 H,s), 1.47(3 H,s), 1.39(3 H,s).

Compound 10b

¹ H NMR (CDCl₃) δ:8.80 ppm (1 H,s), 5.88(1 H,d,J=7.9 Hz), 5.09(1 H,t,J=10.0 Hz), 3.95(1 H,dd), 3.75(2 H,m), 3.48(1 H,ddd), 2.05(3 H,s),1.68(3 H,s), 1.39(3 H,s).

Compound 11a

¹ H NMR (CDCl₃) δ:7.43 ppm (1 H,m), 6.76(2 H,m), 5.46(1 H,t), 4.99(1H,d), 4.75(1 H,d), 4.70(1 H,d), 4.51(1 H,t), 3.97(3 H,s) 3.94(3 H,s),3.81(1 H,m), 3.76(1 H,m), 3.10(1 H,dd), 2.10(3 H,s), 1.46(3 H,s), 1.35(3H,s).

Compound 11b

¹ H NMR (CDCl₃) δ:6.95-6.80 ppm (3 H,m), 5.92(1 H,t,J=10.0 Hz), 4.85(1H,d), 4.61(1 H,d), 4.50(1 H,d,J=8.0 Hz), 3.95(1 H,dd), 3.88(1 H,t,J=10.4Hz), 3.80(1 H,t,J=9.3 Hz), 3.63(1 H,t,J=9.0 Hz), 3.46(1 H,dd,J=9.0,10.0Hz), 3.29(1 H,m), 2.11(3 H,s), 1.46(3 H,s), 1.35(3 H,s).

Compound 12

¹ H NMR (CDCl₃) δ:6.98-6.84 ppm (3 H,m), 4.86(1 H,d), 4.62(1 H,d),4.46(1 H,d), 3.96(1 H,dd), 3.90(6 H,s), 3.83(1 H,t), 3.60(1 H,t),3.50-3.40(2 H,m), 3.23(1 H,m), 1.52(3 H,s), 1.42(3 H,s).

Compound 13

¹ H NMR (CDCl₃) δ:6.94-6.82 ppm (3 H,m), 5.93(1 H,m), 5.35(1 H,d,J=17.09Hz), 5.25(1 H,d,J=10.26 Hz), 5.09(1 H,m), 4.92(1 H,t,J=9.53 Hz), 4.84(1H,d,J=11.23 Hz), 4.60(2 H,m), 4.50(1 H,d,J=7.81 Hz), 3.95(1 H,dd,J=5.35,10.99 Hz), 3.88(3 H,s), 3.87(3 H,s), 3.79(1 H,t,J=4.88 Hz),3.62(1 H,t,J=9.77 Hz), 3.21(1 H,m), 2.78(1 H,m), 2.62(1 H,m),1.7-1.56(mH), 1.45(3 H,s), 1.35(3 H,s), 1.27-1.24(mH), 0.86(3 H,t).

Compound 14

¹ H NMR (CDCl₃) δ:6.98-6.82 ppm (3 H,m), 5.91(1 H,m), 5.37(1 H,dd),5.29(1 H,d), 5.10(1 H,m), 4.88-4.79(2 H,m), 4.68-4.60(2 H,m), 4.50(1H,d), 3.94(1 H,m), 3.89(3 H,s), 3.88(3 H,s), 3.84(1 H,m), 3.64(1 H,m),3.48-3.35(2 H,m), 2.75-2.62(2 H,m), 1.80-1.62(2 H,m), 1.42-1.20(mH),0.86(3 H,t).

Compound 15

¹ H NMR (CDCl₃) δ:6.95-6.81 ppm (3 H,m), 5.90(1 H,m), 5.35(1H,dd,J=1.46,17.33 Hz), 5.30(1 H,dd,J=1.22,10.49 Hz), 5.08(1 H,m), 4.86(1H,d,J=11.47 Hz), 4.82(1 H,t,J=9.1 Hz), 4.60(3 H,m), 4.43(1 H,d,J=7.65Hz), 3.91(1 H,m), 3.88(3 H,s), 3.87(3 H,s), 3.52(1 H,dt,J=2.0,9.28,9.28Hz), 3.42(1 H,dd,J=8.05,10.5 Hz), 3.35(1 H,m), 2.75(1 H,dd,J=7.57,15.39Hz), 2.66(1 H,dd,J=4.88,15.38 Hz), 1.78-1.62(mH), 1.40-1.20(mH), 0.91(9H,s), 0.86(3 H,t), 0.10(3 H,s), 0.09(3 H,s).

Compound 16

¹ H NMR (CDCl₃) δ:6.94-6.82 ppm (3 H,m), 5.99-5.84(2 H,m), 5.33(2 H,m),5.26(2 H,m), 5.06(1 H,m), 5.00(1 H,t), 5.88(1 H,d), 5.81(1 H,t),4.66-4.56(5 H,m), 4.44(1 H,d,J=8.05 Hz), 3.88(3 H,s), 3.87(3 H,s),3.80(1 H,m), 3.56-3.48(2 H,m), 2.71-2.59(2 H,m), 1.65(mH),1.49-1.29(mH), 0.90(9 H,s), 0.89-0.86(3 H,m), 0.86(6 H,s).

Compound 17

¹ H NMR (CDCl₃) δ:6.95-6.82 ppm (3 H,m), 5.95-5.86(2 H,m), 5.36(2 H,d),5.25(2 H,t), 5.04(2 H,m), 4.86(2 H,m), 4.62(5 H,m), 4.50(1 H,d,J=8.05Hz), 3.88(3 H,s), 3.87(3 H,s), 3.84(1 H,m), 3.67(1 H,dd,J=4.15,12.7 Hz),3.48(1 H,m), 2.72-2.60(2 H,m), 1.73-1.50(mH), 1.40-1.28(mH), 0.91-0.82(3H,m).

Compound 18

¹ H NMR (CDCl₃) δ:6.96-6.80 ppm (3 H,m), 5.39(1 H,m), 5.30(1 H,m),5.20(1 H,m), 4.90(1 H,t), 4.84(1 H,d), 4.61(1 H,d), 4.51(1 H,d), 3.96(1H,dd), 3.89(3 H,s), 3.88(3 H,s), 3.80(1 H,t), 3.65(1 H,t), 3.46(1 H,dd),3.29(1 H,m), 2.69(1 H,dd), 2.58(1 H,dd), 228(1 H,t), 2.10-1.96(2 H,m),1.72-1.58(2 H,m), 1.42(3 H,s), 1.35(3 H,s), 1.32-1.18(mH), 0.89-0.82(6H,m).

Compound 19

¹ H NMR (CDCl₃) δ:6.98-6.82 ppm (3 H,m), 5.42(1 H,m), 5.30(1 H,m),5.12(1 H,m), 4.88(1 H,d), 4.82(1 H,t), 4.65(1 H,d), 4.49(1 H,d), 3.94(1H,m), 3.90(3 H,s), 3.89(3 H,s), 3.83(1 H,m), 3.60(1 H,t), 3.45-3.35(2H,m), 2.30(2 H,t), 2.09-1.98(4 H,m), 1.67(mH), 1.40-1.22(mH),0.91-0.85(6 H,m).

Compound 20

¹ H NMR (CDCl₃) δ:6.94-6.81 ppm (3 H,m), 5.94(1 H,m), 5.39-5.26(4 H,m),5.13(1 H,m), 4.86(1 H,d), 4.79(1 H,m), 4.68-4.59(mH), 4.52(1 H,d),4.42(2 H,m), 3.88(3 H,s), 3.87(3 H,s), 3.52(2 H,m), 3.45(1 H,t,J=8.06Hz), 2.60(2 H,m), 2.30(2 H,t), 2.10-1.95(4 H,m), 1.70-1.53(mH),1.40-1.27(mH), 0.88-0.84(6 H,m).

Compound 21

¹ H NMR (CDCl₃) δ:6.94-6.80 ppm (3 H,m), 5.97-5.83(3 H,m), 5.40-5.22(8H,m), 4.96(1 H,dd,J=9.28,10.26 Hz), 4.84(1 H,d,J=11.48 Hz), 4.68-4.29(11H,m), 3.87(3 H,s), 3.86(3 H,s), 3.60(1 H,m), 3.49(1 H,dd,J=8.06,10.26Hz), 2.68(2 H,t), 2.26(2 H,q), 2.08-1.90(4 H,m), 1.70-1.65(mH),1.32-1.18(mH), 0.84(6 H,m).

Compound 22

¹ H NMR (CDCl₃) δ:5.95-5.84 ppm (3 H,m), 5.59(2 H₃β,t), 5.40-5.21(9H,m), 5.03(1 H₃α,t), 4.76-4.22(10 H,m), 3.42(1 H₂β,dd,J=8.05,10.5 Hz),3.20(1 H₂α,dd,J=3.17,10.5 Hz), 2.76-2.62(2 H,m), 2.30-2.23(2 H,m),2.09-1.95(4 H,m), 1.70-1.65(mH), 1.35-1.18(mH), 0.90-0.80(6 H,m).

Compound 23A

¹ H NMR (CDCl₃) δ:8.85 ppm (1 H,s), 6.45(1 H,d,J=3.66 Hz), 5.96-5.84(3H,m), 5.57(1 H,dd,J=9.03,10.74 Hz), 5.41-5.22(9 H,m), 4.60-4.42(8 H,m),4.32(1 H,dd,J=3.91,11.97 Hz), 4.18(1 H,d,J=8.79 Hz), 3.58(1H,dd,J=3.0,10.0 Hz), 2.80-2.67(2 H,m), 2.29(2 H,t), 2.06-1.95(4 H,m),1.70-1.59(mH), 1.35-1.25(mH), 1.81-1.90(6 H,m).

Compound 23B

¹ H NMR (CDCl₃) δ:8.80 ppm (1 H,s), 5.98-5.82(3 H,m), 5.72(1 H,d),5.42-5.22(9 H,m), 5.15(1 H,t), 4.62-4.41(8 H,m), 4.23(2 H,m), 3.72(1H,t), 2.81-2.68(2 H,m), 2.30(2 H,m), 2.10-1.96(4 H,m), 1.71-1.52(mH),1.37-1.15(mH), 0.88(6 H,m).

Compound 24

¹ H NMR (CDCl₃) δ:5.95-5.86 ppm (3 H,m), 5.99-5.83(5 H,m), 5.40-5.22(12H,m), 5.10-5.22(2 H,m), 4.91(1 H,d,J=10.7 Hz), 4.68(1 H,dd),4.64-4.43(10 H,m), 4.33(1 H,q), 4.27(1 H,dd,J=4.9,12.8 Hz), 4.00(1H,d,J=9.9 Hz), 3.89(3 H,s), 3.88(3 H,s), 3.75(2 H,m), 3.60(1 H,m),3.54(1 H,dd,J=7.9,9.8 Hz), 3.47(1 H,dd,J=8.6,11.0 Hz), 2.78-2.58(4 H,m),2.30-2.23(2 H,t), 2.03-1.95(4 H,m), 1.70-1.59(2 H,m), 1.39-1.25(mH),0.90-0.82(9 H,m).

Compound 25

¹ H NMR (CDCl₃) δ:7.23 ppm (1 H,m), 6.92-6.82(2 H,m), 5.98-5.85(5 H,m),5.40-5.21(12 H,m), 5.08-4.99(2 H,m), 4.85(1 H,m), 4.62-4.42(10 H,m),4.38-4.23(2 H,m), 3.89(6 H,s), 3.75-3.61(2 H,m), 2.96(1 H,dd), 2.72(1H,dd), 2.61(2 H,m), 2.29-2.15(2 H,m), 2.02-1.94(4 H,m), 1.70-1.52(mH),1.38-1.20(mH), 0.91-0.82(9 H,m).

Compound 26

¹ H NMR (CDCl₃) δ:7.29 ppm (2 H,d), 7.16(2 H,d), 6.91-6.85(7 H,m),6.40(1 H,d), 6.08(1 H,d), 5.98-5.85(5 H,m), 5.40-5.15(13 H,m), 5.07(1H,t), 4.98(1 H,m), 4.76-4.23(mH), 3.98(1 H,q), 3.84(3 H,s), 3.83(2 H,m),3.82(3 H,s), 3.79(3 H,s), 3.77(3 H,s), 3.82(3 H,m), 3.60-3.50(2 H,m),2.70-2.42(4 H,m), 2.36-2.22(4 H,m), 2.07-1.96(2 H,m), 1.68-1.36(mH),1.33-1.15(mH), 0.91-0.82(15 H,m).

Compound 27

¹ H NMR (CDCl₃) δ:6.35 ppm (1 H,d), 6.12(1 H,d), 5.98-5.82(5 H,m),5.44-5.21(13 H,m), 5.00(1 H,m), 4.94(1 H,d), 4.68-4.48(mH), 4.32(1H,dd), 4.26-4.15(2 H,m), 4.00(1 H,m), 3.92-3.82(2 H,m), 3.79-3.65(2H,m), 3.58(1 H,dd), 2.68-2.49(4 H,m), 2.41-2.22(6 H,m), 2.09-1.97(4H,m), 1.70-1.19(mH), 0.91-0.80(15 H,m).

Compound 28

¹ H NMR (CDCl₃) δ:6.80 ppm (1 H,d,J=8.2 Hz), 6.52(1 H,d,J=7.6 Hz),6.01-5.84(5 H,m), 5.72(1 H,dd,J=2.7,5.4 Hz), 5.45-5.17(13 H,m),5.02-4.90(2 H,m), 4.94(1 H,d,J=8.1 Hz), 4.67-4.33(mH), 4.39-4.28(3 H,m),4.12(1 H,m), 3.99-3.85(3 H,m), 3.89-3.68(4 H,m), 2.64-2.52(4 H,m),2.36-2.12(6 H,m), 2.10-1.95(4 H,m), 1.70-1.15(mH), 0.91-0.81(15 H,m).

Compound 29

¹ H NMR (CDCl₃) δ:7.30 ppm (1 H,d), 6.55(1 H,d), 6.00-5.85(5 H,m),5.78(1 H,dd), 5.44-5.19(13 H,m), 5.10-4.82(3 H,m), 4.67-4.46(mH),4.38-4.28(2 H,m), 4.15(1 H,m), 4.00-3.86(2 H,m), 3.86-3.60(mH),3.40-3.34(4 H,m), 2.66-2.42(6 H,m), 2.37-2.22(6 H,m), 2.08-1.96(4 H,m),1.69-1.20(mH), 0.87 (15 H,m).

Compound 30

¹ H NMR (CDCl₃) δ:7.38 ppm (1 H,d), 7.09(1 H,d), 6.00-5.83(5 H,m),5.71(1 H,m), 5.43-5.19(13 H,m), 5.01(1 H,m), 4.87(2 H,m), 4.68-4.44(mH),4.40-4.36(mH), 4.18(1 H,m), 3.93(1 H,dd), 3.76(1 H,q), 3.71-3.62(2 H,m),3.38(2 H,q), 3.32(2 H,q), 2.68-2.41(8 H,m), 2.28-2.20(2 H,t),2.07-1.95(4 H,m), 1.68-1.48(mH), 1.35-1.12(mH), 0.90-0.81(15 H,m).

Analog B214 (Compound 31)

R_(f) :0.43 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v)] RT(HPLC) 12.22 min

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ:5.40 ppm (1 H,m), 5.17(1 H,m),5.09(1 H,m), 5.00(2 H,m), 4.96(1 H,t,J=10.0 Hz), 4.48(1 H,d), 4.02(2H,m), 3.90-3.60(mH), 3.45(1 H,m), 3.34(1 H,t,J=9.6 Hz), 3.26-3.13(mH),2.48-2.20(mH), 2.06(2 H,t), 1.84(4 H,m), 1.45-1.00(mH), 0.65(15 H,m).

¹³ C NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ:205.26 ppm 205.16, 173.71, 172.57,170.52, 168.10, 167.45, 130.63, 127.93, 100.63, 94.55, 74.64, 72.74,72.42, 71.60, 69.82, 67.95, 67.64, 59.95, 53.57, 43.03, 42.79, 41.74,38.26, 36.78, 34.63, 33.91, 33.47, 31.51, 31.46, 31.43, 31.38, 29.25,29.21, 29.15, 29.12, 29.08, 29.06, 28.96, 28.93, 28.86, 28.74, 28.65,28.56, 26.79, 26.12, 25.15, 24.78, 24.53, 23.00, 22.91, 22.24, 22.22,13.51.

³¹ P NMR (CDCl₃ :CD₃ OD 3:1, v/v) δ:1.31 ppm, -1.40.

Compound 32

¹ H NMR (CDCl₃) δ:6.89-6.80 ppm (3 H,m), 6.68(1 H,d,J=8.79 Hz), 6.56(1H,d,J=8.05 Hz), 5.95-5.86(5 H,m), 5.41-5.28(13 H,m), 4.97(1 H,m), 4.80(1H,t,J=9.7 Hz), 4.72(1 H,d,J=8.5 Hz), 4.60(mH), 4.48(2 H,m), 4.30(2 H,m),3.97(2 H,t), 3.89(3 H,s), 3.85(3 H,s), 3.77-3.74(2 H,m), 3.68-3.60(2H,m), 3.00-2.56 (mH), 2.26(2 H,t), 2.09-1.04(mH), 0.88-0.84(15 H,m).

Compound 33

¹ H NMR (CDCl₃) δ:6.83 ppm (1 H,d,J=7.33 Hz), 6.59(1 H,d,J=9.03 Hz),5.94-5.86(5 H,m), 5.47(1 H,t,J=7.1 Hz), 5.40-5.30(12 H,m), 5.00(1 H,m),4.68-4.45(mH), 4.38(1 H,q), 4.32-4.20(2 H,m), 3.83-3.69(2 H,m), 3.41(1H,q), 2.98-2.56(8 H,m), 2.27(2 H,t), 2.07-1.90(mH), 1.67-1.52(mH),1.35-1.15(mH), 0.89-0.85(15 H,m).

Compound 34

¹ H NMR (CDCl₃) δ:7.39 ppm (1 H,d), 7.08(1 H,d), 6.00-5.83(7 H,m),5.70(1 H,dd), 5.43-5.20(13 H,m), 5.01(1 H,m), 4.88(1 H,m),4.65-4.45(mH), 4.40-4.25(2 H,m), 4.18(1 H,m), 3.92(1 H,dd), 3.75(1 H,q),3.67(1 H,m), 3.39(2 H,d), 3.32(2 H,q), 2.67-2.42(4 H,m), 2.24(2 H,t),2.07-1.92(2 H,m), 1.74-1.46(mH), 1.35-1.24(mH), 0.91-0.82(15 H,m).

Compound 36

R_(f) :0.77 [hexanes:ethyl acetate, 1:1 (v/v)]

¹ H NMR (CDCl₃) δ:4.92 ppm (1 H,t,J=9.0 Hz), 4.62(1 H,d, J=7.9 Hz),3.88(1 H,dd), 3.77(1 H,t), 3.64(1 H,t), 3.30(2 H,m), 2.21(3 H,s), 1.44(3H,s), 1.35(3 H,s), 0.89(9 H,s), 0.12(3 H,s). 0.11(3 H,s).

Compound 37

R_(f) :0.37 [hexanes:ethyl acetate, 3:1 (v/v)]

¹ H NMR (CDCl₃) δ:4.70 ppm (1 H,d,J=7.5 Hz), 3.86 (1 H,dd,J=3.0,8.35Hz), 3.77(1 H,t,J=10.3 Hz)3.57(1 H,t,J=9.29 Hz), 3.45(1 H,m), 3.30(2H,m), 1.50(3 H,s), 1.42(3 H,s), 0.91(9 H,s), 0.14(3 H,s), 0.13(3 H,s).

Compound 38

¹ H NMR (CDCl₃) δ:5.90 ppm (1 H,m), 5.35(1 H,dd,J=1.46,17.36 Hz), 5.23(1H,d,J=10.49 Hz), 5.08(1 H,m), 4.90(1 H,t,J=10.01 Hz), 4.60(3 H,m),3.86(1 H,dd, J=5.61,10.98 Hz), 3.74(1 H,t,J=10.5 Hz), 3.62(1 H,t,J=9.52Hz), 3.30(2 H,m), 2.75(1 H,dd, J=7.0, 15.8 Hz), 2.63(1 H,dd,J=6.35,15.63Hz), 1.65(mH), 1.42(3 H,s), 1.32(3 H,s), 1.24(mH), 0.89(9 H,s), 0.86(3H,t), 0.12(3 H,s), 0.10(3 H,s).

Compound 39

¹ H NMR (CDCl₃) δ:5.90 ppm (1 H,m), 5.38(1 H,dd), 5.29(1 H,d), 5.07(1H,m), 4.81(1 H,t), 4.62(2 H,m), 3.88(1 H,m), 3.80(1 H,m), 3.62(1 H,m),3.40(1 H,m), 3.31(1 H,dd), 2.79(2 H,m), 1.98(1 H,t), 1.72(1 H,m), 1.65(1H,m), 1.27(mH), 0.91(9 H,s), 0.88(3 H,t), 0.18(3 H,s), 0.17(3 H,s).

Compound 40

¹ H NMR (CDCl₃) δ:5.96-5.85 ppm (4 H,m), 5.42-5.22(11 H,m), 5.05(1 H,m),5.00(1 H,t), 4.80(1 H,dd), 4.66-4.42(mH), 4.35-4.22(2 H,m), 4.10(1 H,d),3.61(1 H,m), 3.52(1 H,m), 3.42(1 H,dd), 3.31(1 H,dd), 2.78-2.62(4 H,m),2.28(2 H,t), 2.09-1.95(4 H,m), 1.69-1.87(4 H,m), 1.38-1.19(mH), 0.92(9H,s), 0.85(15 H,m), 0.16(6 H,s).

Compound 41

¹ H NMR (CDCl₃) δ:5.94-5.86 ppm (5 H,m), 5.38-5.21(12 H,m), 5.19-4.97(2H,m), 4.72-4.42(mH), 4.38(1 H,d,J=8.06 Hz), 4.32-4.23(2 H,m), 3.88(1H,m), 3.75(1 H,dd), 3.60(1 H,m), 3.40(2 H,m), 2.76(6 H,m), 2.28(2 H,t),2.09-1.95(4 H,m), 1.70-1.63(4 H,m), 1.35-1.19(mH), 0.92(9 H,s),0.87-0.86(15 H,m), 0.17(3 H,s), 0.16(3 H,s).

Compound 43

¹ H NMR (CDCl₃) δ:7.20 ppm (1 H,d), 7.00(1 H,d), 5.95(5 H,m), 5.30(mH),4.95(1 H,q), 4.75(1 H,t), 4.55(mH), 4.30(mH), 3.90(1 H,dd), 3.70(mH),3.45(mH), 3.35(2 H,s), 3.28(2 H,s), 2.60(1 H,dd), 2.45(mH), 2.25(2 H,t),1.95(mH), 1.65(mH), 1.50(mH), 1.20(mH), 0.85(15 H,m), 0.80(9 H,s),0.08(6 H,2 s).

Compound 44

¹ H NMR (CDCl₃) δ:7.20 ppm (1 H,d), 6.82(1 H,d), 5.95(5 H,m), 5.30(mH),5.10(mH), 4.58(mH), 4.47(mH), 4.30(mH), 3.70(mH), 3.35(mH), 2.50(mH),2.25(2 H,t), 2.00(mH), 1.50(mH), 1.25(mH), 0.85(15 H,m).

Compound 45

R_(f) :0.53 [hexanes:ethyl acetate, 4:1 (v/v)]

¹ H NMR (CDCl₃) δ:4.53 ppm (1 H,d,J=7.42 Hz), 3.99(1 H,m), 3.87-3.73(2H,m), 3.60(1 H,t,J=9.2 Hz), 3.26-3.14(2 H,m), 1.70-1.63(2 H,m), 1.48(3H,s), 1.40(3 H,s), 1.27(mH,br.s), 0.91(9 H,s), 0.90-0.85(3 H,m), 0.13(3H,s), 0.21(3 H,s).

Compound 46

R_(f) :0.80 [hexanes:ethyl acetate, 4:1 (v/v)]

¹ H NMR (CDCl₃) δ:5.43-5.28 ppm (2 H,m), 5.03(1 H,m), 4.49(1 H,d,J=7.46Hz), 3.86-3.73(2 H,m), 3.66-3.56(2 H,m), 3.22-3.10(2 H,m), 2.30-2.26(2H,t), 2.09-1.97(mH), 1.83-1.54(mH), 1.48(3 H,s), 1.38(3 H,s),1.26(mH,br.s), 0.91(9 H,s), 0.89-0.85(6 H,m), 0.13(3 H,s), 0.12(3 H,s).

Compound 47

R_(f) :0.13 [hexanes:ethyl acetate, 4:1 (v/v)]

¹ H NMR (CDCl₃) δ:5.44-5.28 ppm (2 H,m), 5.10-5.04(1 H,m), 4.53(1H,d,J=7.6 Hz), 3.91-3.85(2 H,m), 3.77-3.66(2 H,m), 3.43(1 H,m), 3.33(1H,m), 3.18(1 H,dd,J=6,9.9 Hz), 3.01(1 H,dd,J=9.3,9.8 Hz), 2.32-2.26(2H,t), 2.10-1.48(mH), 1.34-1.25(mH), 0.92(9 H,s), 0.92-0.85(6 H,m),0.15(3 H,s), 0.14(3 H,s).

Compound 48

R_(f) :0.45 [hexanes:ethyl acetate, 4:1 (v/v)]

¹ H NMR (CDCl₃) δ:5.95 ppm (1 H,m), 5.33(2 H,m), 5.30(2 H,m), 5.07(1H,m), 4.62(2 H,d), 4.48(1 H,d,J=7.8 Hz), 4.44(1 H,dd,J=2.2,11.3 Hz),4.33(1 H,dd,J=6.1,11.7 Hz), 3.85(1 H,m), 3.68(1 H,m), 3.58(1H_(OH),d,J=3.2 Hz) 3.45(1 H,m), 3.37(1 H,m), 3.18(1 H,t,J=9.1 Hz),2.98(1 H,t,J=10.1 Hz), 2.28(2 H,t), 2.06(mH), 1.82(mH), 1.65(mH),1.25(mH), 0.91(9 H,s), 0.85(6 H,m), 0.13(6 H,2 s).

Compound 49

R_(f) :0.29 [hexanes:ethyl acetate, 4:1 (v/v)]

¹ H NMR (CDCl₃) δ:5.95 ppm (3 H,m), 5.41-5.26(8 H,m), 5.00(1 H,m),4.63-4.47(mH), 4.30(1 H,dd,J=6.6,11.7 Hz), 4.18(1 H,q), 3.72(1 H,m),3.55(1 H,m), 3.25(2 H,t,J=7.9,10.3 Hz), 3.15(1 H,t,J=8.8,10.8 Hz),2.25(2 H,t), 2.00(4 H,m), 1.65(mH), 1.50(mH), 1.25(mH), 0.91(9 H,s),0.85(6 H,m), 0.01(6 H,2 s).

Compound 50

R_(f) :0.25 & 0.20 [diethyl ether:dichloromethane, 1:9 (v/v)]

¹ H NMR (CDCl₃) δ:5.95 ppm (3 H,m), 5.35(mH), 5.25(mH), 4.95(mH),4.59(mH), 4.30(mH), 3.75(mH), 3.60(mH), 3.35(mH), 2.25(2 H,t), 2.00(mH),1.65(mH), 1.25(mH), 0.85(6 H,m).

Compound 51A

R_(f) :0.50 [diethyl ether:dichloromethane, 1:9 (v/v)]

¹ H NMR (CDCl₃) δ:8.77 ppm (1 H,s), 6.38(1 H,d,J=2.41 Hz), 5.95(3 H,m),5.30(mH), 4.98(1 H,m), 4.55(mH), 4.40(mH), 4.05(1 H,m), 3.85(1 H,m),3.75(1 H,m), 3.60(1 H,dd,J=3.42,9.5 Hz), 2.27(2 H,t), 2.00(mH),1.65(mH), 1.25(mH), 0.85(6 H,m).

Compound 51B

Rf:0.37 [diethyl ether:dichloromethane, 1:9 (v/v)]

¹ H NMR (CDCl₃) δ:8.80 ppm (1 H,s), 5.95(3 H,s), 5.95(3 H,m), 5.72(1H,d), 5.30(mH), 5.15(1 H,t), 4.62(1 H,d), 4.50(mH), 4.32(1 H,dd), 3.80(1H,m), 3.70(1 H,t), 2.70(2 H,t), 2.25(2 H,t), 1.95(mH), 1.60(mH),1.25(mH), 0.85(6 H,m).

Compound 52

R_(f) :0.38 [ethyl acetate:hexanes, 1:9 (v/v)]

¹ H NMR (CDCl₃) δ:5.98-5.88 ppm (1 H,m), 5.32(1 H,m), 5.25(1 H,m),4.88(1 H,m), 4.62(2 H,d), 4.48(1 H,d,J=7.46 Hz), 3.86-3.66(mH), 3.59(1H,t,J=9.4 Hz), 3.23-3.10(mH), 1.87-1.80(2 H,m), 1.64-1.56(2 H,m), 1.48(3H,s), 1.38(3 H,s), 0.91(9 H,s), 0.90-0.86(3 H,t), 0.13(3 H,s), 0.12(3H,s).

Compound 53

R_(f) :0.12 [ethyl acetate:hexanes, 1:4 (v/v)]

¹ H NMR (CDCl₃) δ:5.93 ppm (1 H,m), 5.37(1 H,m), 5.27(1 H,m), 4.90(1H,m), 4.67-4.58(2 H,m), 4.54(1 H,d,J=7.5 Hz), 4.12(1 H,q), 3.94(1 H,m),3.87(1 H,dd,J=3.4,11.5 Hz), 3.78-3.69(2 H,m), 3.50-3.43(2 H,m),3.34-3.30(1 H,m), 3.21(1 H,dd,J=7.6,10.0 Hz), 3.03(1 H,t,J=9.2 Hz),2.05-2.01(1 H,m), 1.96-1.50(mH), 1.36-1.23(mH), 0.92(9 H,s), 0.88-0.85(3H,m), 0.15(3 H,s), 0.14(3 H,s).

Compound 54

¹ H NMR (CDCl₃) δ:5.91 ppm (1 H,m), 5.37-5.32(1 H,m), 5.26-5.23(1 H,m),4.92-4.87(1 H,m), 4.65-4.55(2 H,m), 4.47(1 H,d,J=7.64 Hz),3.90-3.74(mH), 3.48(1 H,ddd,J=2.1,9.3,11.2 Hz), 3.42(1 H_(OH),d,J=2.1Hz), 3.28(1 H,m), 3.18(1 H,dd,J=7.6,9.9 Hz), 3.00(1 H,dd,J=8.7,9.8 Hz),1.89(mH), 1.60-1.20(mH), 0.91(18 H,s), 0.90-0.84(3 H,m), 0.065(6 H,s),0.059(6 H,s).

Compound 55

¹ H NMR (CDCl₃) δ:5.97-5.87 ppm (2 H,m), 5.37-5.31(2 H,m), 5.28-5.23(2H,m), 4.81(1 H,m), 4.69(1 H,t,J=10.0 Hz), 4.68-4.57(2 H,m), 4.48(1H,d,J=7.5 Hz), 3.79(1 H,q), 3.75-3.65(2 H,d), 3.64-3.58(1 H,m), 3.36(1H,m), 3.27(1 H,dd,J=7.7,10.1 Hz), 3.17(1 H,t,J=9.3 Hz), 1.80(2 H,q),1.61-1.52(mH), 1.31-1.25(mH), 0.91(18 H,m), 0.92-0.84(3 H,m), 0.025(6H,s), 0.010(6 H,s).

Compound 56

¹ H NMR (CDCl₃) δ:5.96-5.97 ppm (2 H,m), 5.38-5.20(4 H,m), 4.80(1 H,m),4.69-4.56(mH), 4.52(1 H,d,J=7.4 Hz), 3.81-3.59(3 H,m), 3.36(1 H,m),3.30-3.26(1 H,dd,J=7.64,9.88 Hz), 3.20(1 H,t,J=9.25 Hz), 2.19(1H_(OH),t,J=5.71 Hz), 1.82(2 H,q), 1.68-1.50(mH), 1.34(mH,br.s), 0.92(9H,s), 0.85(3 H,m), 0.14(6 H,s).

Compound 57A

¹ H NMR (CDCl₃) δ:5.43-5.28 ppm (2 H,m), 5.06(1 H,m), 4.47(1 H,d, J=7.68Hz), 3.83(1 H,m), 3.66-3.56(2 H,m), 3.47(1 H,m), 3.38(3 H,s), 3.34(1H,m), 3.20(1 H,t,J=7.9 Hz), 2.98(1 H,t,J=8.9 Hz), 2.29(2 H,t),2.09-1.50(mH), 1.34-1.25(mH), 0.92(9 H,s), 0.91-0.85(6 H,m), 0.14(6H,s).

Compound 57B

¹ H NMR (CDCl₃) δ:7.89-7.34 ppm (10 H,m), 5.43-5.29(2 H,m), 5.00(1 H,m),4.44(1 H,d,J=7.74 Hz), 3.84-3.70(3 H,m), 3.46(3 H,s), 3.28(1 H,q),3.18(2 H,m), 3.04(1 H,t,J=9.5 Hz), 2.30(2 H,t), 2.11-2.00(4 H,m), 1.89(2H,m), 1.70(2 H,m), 1.58(2 H,m), 1.27(mH), 1.07(9 H,s), 0.95(9 H,s),0.87(6 H,m), 0.16(3 H,s), 0.15(3 H,s).

Compound 58

¹ H NMR (CDCl₃) δ:5.96-5.87 ppm (2 H,m), 5.41-5.23(mH), 5.00(1 H,m),4.58(4 H,m), 4.46(1 H,d,J=7.50 Hz), 4.23(1 H,q), 3.77-3.68(mH), 3.57(1H,dd,J=5.13,10.91 Hz), 3.41(1 H,m), 3.35(3 H,s), 3.24(1 H,dd, J=7.7,10.1Hz), 3.12(1 H,dd,J=8.8,9.7 Hz), 2.28-2.26(2 H,t), 2.07-1.52(mH),1.32-1.12(mH), 0.91(9 H,s), 0.90-0.85(6 H,m), 0.13(6 H,s).

Compound 59

¹ H NMR (CDCl₃) δ:5.97-5.90 ppm (2 H,m), 5.41-5.25(4 H,m), 4.97(1 H,m),4.61-4.54(4 H,m), 4.27-4.07(2 H,m), 3.81-3.40(mH), 3.38(3 H,s), 3.32(2H,m), 3.22(1 H,t,J=9.9 Hz), 2.29-2.24(2 H,m), 2.08-1.52(mH),1.33-1.23(mH), 0.88-0.84(6 H,m).

Compound 60A and B

R_(f)α/β :0.53 [hexanes:ethyl acetate, 1:1 (v/v)]

¹ H NMR (CDCl₃) δ:8.74 ppm (H1.sub.α,β,2 s), 6.39(1 H.sub.α,d,,J=3.46Hz), 6.00-5.91(2 H,m), 5.58(1 H.sub.β,d,J=8.46 Hz), 5.42-5.25(mH),5.00(1 H,m), 4.63-4.36(mH), 3.99-3.50(mH), 3.37(3 H.sub.β,s), 3.36(3H.sub.α,s), 3.34-3.30(mH), 2.35-2.25(2 H,m), 2.08-1.80(mH),1.70-1.54(mH), 1.27-1.23(mH), 0.89-0.85(6 H,m).

Compound 62

¹ H NMR (CDCl₃) δ:6.96 ppm (1 H,s), 6.85(2 H,m), 5.95(5 H,m), 5.30(mH),5.00(mH), 4.35(1 H,d), 4.27(1 H,dd), 4.20(1 H,,q), 3.98(1 H,d), 3.85(6H,2 s), 3.75(mH), 3.52(2 H,m), 3.35(1 H,t), 3.15(1 H,t), 2.60(2 H,m),2.25(2 H,t), 2.00(4 H,m), 1.60(mH), 1.25(mH), 0.85(9 H,m).

Compound 63

¹ H NMR (CDCl₃) δ:6.98 ppm (1 H,d,J=1.71 Hz), 6.90(1 H,d,J=8.30 Hz),6.85(1 H,d,J=8.30 Hz), 5.95(4 H,m), 5.30(mH), 5.00(1 H,t,J=9.03 Hz),4.90(mH), 4.55(mH), 4.30(mH), 3.85(6 H,2 s), 3.70(mH), 3.62(1 H,m),3.50(mH), 3.35(mH), 3.00(1 H,t,J=9.8 Hz), 2.69(2 H,t,), 2.25(2 H,t),2.00(mH), 1.60(mH), 1.25(mH), 0.85(9 H,m).

Compound 64

¹ H NMR (CDCl₃) δ:5.95 ppm (4 H,m), 5.40-5.22(10 H,m), 4.96(1 H,m),4.90(1 H,m), 4.55(mH), 4.32(1 H,d), 4.30-4.22(2 H,m), 4.18(1 H,q),4.10(1 H,dd), 3.90(mH), 3.75(mH), 3.50(mH), 3.33(2 H,m), 3.20(mH),3.00(1 H,t), 2.25(2 H,t), 2.00(4 H,m), 1.80(mH), 1.50(mH), 1.25(mH),0.91(9 H,s), 0.85(9 H,m), 0.15(6 H,s).

Compound 65

¹ H NMR (CDCl₃) δ:5.98-5.87 ppm (4 H,m), 5.42-5.23(mH), 4.95(1 H,m),4.80(2 H,m), 4.65-4.50(mH), 4.49(1 H,d,J=7.6 Hz), 4.27(1 H,d,J=8.1 Hz),4.26(1 H,m), 3.87(1 H,d,J=9.95 Hz), 3.83-3.54(mH), 3.36(3 H,s),3.30-3.13(4 H,m), 2.28-2.23(2 H,t), 2.07-1.78(mH), 1.73-1.53(mH),1.40-1.23(mH), 0.92(9 H,s), 0.91-0.77(9 H,m), 0.17(6 H,s).

Compound 67

¹ H NMR (CDCl₃) δ:7.32 ppm (1 H,d,J=8.2 Hz), 7.2491 H,d,J=9.8 Hz),5.97-5.89(4 H,m), 5.37-5.23(mH), 4.91(2 H,m(, 4.81(1 H,m), 4.71(1 H,m),4.63-4.54(mH), 4.24(1 H,q), 3.88-3.43(mH), 3.39(3 H,s), 2.53-2.50(4H,m), 2.26-2.23(2 H,t), 2.06-1.54(mH), 1.2(mH), 0.88-0.83(mH), 0.08(3H,s), 0.05(3 H,s).

Compound 68

R_(f) :0.52 [dichloromethane:methyl alcohol, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ:7.25 ppm (1 H,d,J=7.6 Hz), 7.22(1 H,d,J=8.4 Hz),5.97-5.85(4 H,m), 5.41-5.20(mH), 5.05(1 H,d,J=8.0 Hz), 4.93(1 H,m),4.78(1 H,m), 4.65-4.51(mH), 4.28(1 H,q), 4.11(1 H,m), 3.79-3.57(mH),3.52-3.39(mH), 3.37(3 H,s), 2.50(4 H,t), 2.25(2 H,t), 2.07-1.96(4 H,m),1.78-1.48(mH), 1.24(mH), 0.86(15 H,m).

Compound 69

¹ H NMR (CDCl₃) δ:7.55 ppm (1 H,d), 7.05(1 H,d), 6.00-5.86(6 H,m),5.79(1 H,m), 5.42-5.20(mH), 4.91(1 H,m), 4.84(2 H,m), 4.75(1 H,t),4.67-4.52(mH), 4.28(1 H,q), 4.13(1 H,m), 4.05(1 H,m), 3.91(1 H,m),3.91(1 H,d), 3.80-3.40(mH), 3.39(3 H,s), 2.52(4 H,m), 2.26(2 H,t),2.10-1.95(4 H,m), 1.82-1.43(mH), 1.38-1.24(mH), 0.87(15 H,m).

Analog B531 (Compound 70)

RT(HPLC):13.87 min.

¹ H NMR (CDCl₃ :CD₃ OD,3:1, v/v) δ:5.29 ppm (1 H,dd,J=3.3,6.3 Hz),5.20(1 H,m), 5.10(1 H,m), 4.70(1 H,m), 4.46(1 H,d,J=8.2 Hz), 3.86(2H,m), 3.72-3.30(mH), 3.20(3 H,s), 3.18(1 H,t), 2.38(4 H,m), 2.10 (2H,t), 1.82(4 H,m), 1.72-0.95(mH), 0.68(15 H,t).

¹³ C NMR (CDCl₃ :CD₃ OD,3:1, v/v) δ:205.9 ppm, 174.1, 168.0, 167.6,130.6, 127.8, 100.3, 94.6, 80.1, 78.9, 74.6, 73.6, 72.8, 71.1, 70.6,69.7, 69.2, 69.1, 69.08, 67.3, 58.3, 54.9, 52.2, 52.1, 48.8, 47.5, 43.0,42.9, 37.3, 36.6, 34.4, 34.1, 33.6, 31.4, 31.3, 29.3, 29.2, 29.1, 29.0,28.9, 28.8, 28.7, 28.6, 28.5, 26.7, 26.1, 25.3, 24.8, 24.6, 22.9, 22.8,22.2, 13.4.

³¹ P NMR (CDCl₃ :CD₃ OD,3:1, v/v) δ:-0.58 ppm, -1.24.

Compound 71

¹ H NMR (CDCl₃) δ:7.10 ppm (1 H,d), 6.95(1 H,d), 5.95(4 H,m), 5.25(mH),4.92(1 H,d), 4.82(1 H,d), 4.75(1 H,m), 4.65(mH), 4.50(mH), 4.38(1 H,q),3.85(1 H,m), 3.45(mH), 3.35(3 H,s), 2.80(4 H,m), 2.63(2 H,m), 2.20(2H,t), 1.95(mH), 1.65(mH), 1.20(mH), 0.80(24 H,m), 0.05(6 H,2 s).

Compound 72

¹ H NMR (CDCl₃) δ:7.22 ppm (1 H,d), 6.65(1 H,d), 5.90-94 H,m),5.40-5.23(mH), 4.90(1 H,m), 4.83(1 H,m), 4.70(1 H,d), 4.60-4.50(mH),4.40(1 H,q), 4.10(1 H,m), 4.02(1 H,m), 3.95(1 H,q), 3.80-3.62(mH),3.47(3 H,m), 3.39(3 H,s), 3.36(1 H,t), 2.98-2.83(4 H,m), 2.24(2 H,t),2.00(4 H,m), 1.80-1.20(mH), 0.85(15 H,m).

Compound 73

¹ H NMR (CDCl₃) δ:7.55 ppm (2 H,m), 5.95(6 H,m), 5.80(1 H,m), 5.35(mH),4.90(1 H,m), 4.60(mH), 4.32(1 H,q), 4.15(1 H,m), 3.65(mH), 3.45(1 H,m),3.35(3 H,s), 2.85(4 H,m), 2.25(2 H,t), 2.00(4 H,m), 1.80(mH), 1.25(mH),0.85(15 H,m).

Compound 74

¹ H NMR (CDCl₃) δ:5.95 ppm (5 H,m), 5.60(1 H,t), 5.30(mH), 5.05(3 H,m),4.75(2 H,m), 4.60(mH), 4.50(mH), 4.40(mH), 4.30(mH), 3.90(1 H,m),3.75(mH), 3.47(mH), 3.28(1 H,dd), 2.68(4 H,m), 2.25(2 H,t), 2.00(4 H,m),1.60(mH), 1.25(mH), 0.85(9 H,m).

Compound 75

¹ H NMR (CDCl₃) δ:8.85 ppm (1 H,s), 6.45(1 H,d,J=3.6 Hz), 5.92(5 H,m),5.60(1 H,t,J=10.2 Hz), 5.30(mH), 5.10(mH), 5.03(1 H,dt,J=4.3,10.1,10.1Hz), 4.62(mH), 4.50(mH), 4.30(1 H,q), 4.25(1 H,m), 4.00(1 H,d,J=10.8Hz), 3.75(1 H,dd,J=4.3,11.5 Hz), 3.70(1 H,dd,J=3.6,11.0 Hz), 3.61(1H,m), 3.40(1 H,dd,J=7.9,9.7 Hz), 2.65(4 H,m), 2.25(2 H,t), 2.00(4 H,m),1.60(mH), 1.25(mH), 0.85(9 H,t).

Compound 76

¹ H NMR (CDCl₃) δ:5.98 ppm (7 H,m), 5.75(1 H,t,J=8.2 Hz), 5.35(mH),5.07(1 H,m), 5.04(1 H,dd,J=9.2,10.1 Hz), 4.95(1 H,t,J=9.8 Hz), 4.62(mH),4.50(mH), 4.47(1 H,d,J=7.5 Hz), 4.30(mH), 4.00(2 H,m), 3.70(1H,dd,J=3.9,11.4 Hz), 3.60(1 H,m), 3.42(1 H,dd,J=7.5,9.8 Hz), 2.70(4H,m), 2.25(2 H,t), 2.00(4 H,m), 1.62(mH), 1.30(mH), 0.85(9 H,t).

Compound 77

¹ H NMR (CDCl₃) δ:5.95 ppm (7 H,m), 5.55(1 H,t), 5.35(mH), 5.05(mH),4.90(1 H,t), 4.62(mH), 4.28(mH), 4.20(mH), 4.05(mH), 3.65(mH), 3.35(1H,q), 2.92(1 H,t), 2.70(mH), 2.60(2 H,d), 2.25(2 H,t), 2.00(mH),1.75(mH), 1.60(mH), 1.30(mH), 0.85(9 H,t).

Compound 78

¹ H NMR (CDCl₃) δ:7.36 ppm (1 H,d,J=8.5 Hz), 7.28(1 H,d), 5.90(7 H,m),5.60(1 H,dd,J=10.6,11.6 Hz), 5.30(mH), 5.05(1 H,m), 4.90(2 H,m), 4.75(1H,t,J=9.5 Hz), 4.55(mH), 4.30(mH), 4.08(2 H,m), 3.85(mH), 3.69(mH),2.65(mH), 2.50(2 H,t), 2.45(2 H,t), 2.25(2 H,t), 2.00(mH), 1.60(mH),1.30(mH), 0.85(15 H,m).

Compound 79a

¹ H NMR (CDCl₃) δ:5.94 ppm (1 H,m), 5.45(1 H,d,J=9.5 Hz), 5.35(1H,dd,J=1.47,17.1 Hz), 5.25(1 H,d,J=10.0 Hz), 4.98(1 H,d,J=3.66 Hz),4.97(1 H,m), 4.25(1 H,dd,J=5.2,12.7 Hz), 4.06(1 H,dd,J=1.2,14.7 Hz),3.90-3.60(mH), 3.14(1 H,dd,J=3.4,10.2 Hz), 2.12(3 H,s), 1.45(3 H,s),1.37(3 H,s).

Compound 79b

¹ H NMR (CDCl₃) δ:5.94 ppm (1 H,m), 5.35(1 H,dd,J=1.5,17.1 Hz), 5.25(1H,dd,J=1.2,17.1 Hz), 4.95(2 H,t,J=9.76 Hz), 4.47(1 H,d,J=7.81 Hz),4.40(1 H,dd,J=5.1,11.7 Hz), 4.16(1 H,dd,J=6.5,12.4 Hz), 3.95(1H,dd,J=5.4,10.8 Hz), 3.80(1 H,t,J=10.7 Hz), 3.65(1 H,t,J=9.76 Hz),3.43(1 H,dd,J=8.1,10.0 Hz), 3.27(1 H,m), 2.12(3 H,s), 1.45(3 H,s),1.37(3 H,s).

Compound 80

¹ H NMR (CDCl₃) δ:4.95 ppm (1 H,d,J=3.9 Hz), 4.12(1 H,m), 3.80(mH),3.60(mH), 3.28(1 H,dd,J=3.66,10.0 Hz), 2.62(1 H,d,J=2.44 Hz), 2.22(1H_(OH),t) 1.51(3 H,s), 1.44(3 H,s).

Compound 81

¹ H NMR (CDCl₃) δ:5.92 ppm (1 H,m), 5.44(1 H,t,J=9.52 Hz), 5.37(1H,dd,J=1.46,17.4 Hz), 5.27(1 H,dd,J=1.2,10.5 Hz), 5.11(mH), 5.02(1H,d,J=3.42 Hz), 4.61(2 H,m), 3.85(mH), 3.65(mH), 3.05(1 H,dd,J=3.66,10.5Hz), 2.79(1 H,dd,J=7.08,14.5 Hz), 2.65(1 H,dd,J=6.35,15.4 Hz), 1.65(mH),1.45(3 H,s), 1.36(3 H,s), 1.25(mH), 0.90(12 H,m), 0.08(6 H,2 s).

Compound 82

¹ H NMR (CDCl₃) δ: 5.95 ppm (1H, m), 5.45(1H,t,J=9.3 Hz),5.37(1H,dd,J=1.47,15.8 Hz), 5.27(1H,dd,J=1.22,10.5 Hz), 5.12(1H,m),4.99(1H,d,J=3.66 Hz), 4.62(2H,m), 3.80(mH), 3.65(mH),3.16(1H,dd,J=3.62,10.5 Hz), 2.78(1H,dd,J=7.02,15.4 Hz),2.63(1H,dd,J=6.30,15.6 Hz), 1.65(mH), 1.46(3H,s), 1.37(3H,s), 1.26(mH),0.86(3H,t).

Compound 83

¹ H NMR (CDCl₃) δ: 5.95 ppm (3H,m), 5.41(mH), 5.27(mH), 5.12(1H,m),5.02(1H,d,J=3.4 Hz), 4.55(mH), 4.25(mH), 3.90(mH), 3.65(1H,t,J=8.8 Hz),3.10(1H,dd,J=3.67,10.5 Hz), 2.79(1H,dd,J=5.4,14.4 Hz),2.63(1H,dd,J=6.6,15.6 Hz), 1.65(mH), 1.45(3H,s), 1.36(3H,s), 1.26(mH),0.86(3H,t).

Compound 84

R_(f) :0.6 [methylene chloride:diethyl ether, 4:1 (v/v)]

¹ H NMR (CDCl₃) δ: 5.95 ppm (3H,m), 5.40(mH), 5.25(1H,dd), 5.13(1H,m),4.98(1H,d,J=3.4 Hz), 4.55(mH), 4.25(mH), 3.80(mH), 3.65(1H,t), 3.38(mH),3.13(1H,dd), 2.75(1H,dd), 2.65(1H,dd), 1.68(mH), 1.45(2H,q), 1.25(mH),0.85(12H,m), 0.10(6H,s).

Compound 85

¹ H NMR (CDCl₃) δ: 5.95 ppm (4H,m), 5.55(2H,m), 5.35(8H,m), 5.10(1H,m),5.01(2H,m), 4.95(1H,t), 4.90(1H,t), 4.55(mH), 4.23(2H,m), 3.90(2H,m),3.68(mH), 3.25(1H,dd), 2.68(1H,dd), 2.58(1H,dd), 2.15(2H,q), 1.25(mH),0.85(mH), 0.10(6H,s).

Compound 86

R_(f) :0.80 [methylene chloride:diethyl ether, 4:1 (v/v)]

¹ H NMR (CDCl₃) δ: 7.55 ppm (1H,d,J=8.8 Hz), 5.95(4H,m), 5.30(mH),4.95(mH), 4.85(1H,t,J=10.0 Hz), 4.78(1H,d,J=3.6 Hz), 4.55(mH),4.35(1H,m), 4.25(mH), 3.85(mH), 3.71(2H,d), 3.62(1H,m), 3.20(1H,d,J=13.9Hz), 3.16(1H,m), 2.85(1H,d,J=13.9 Hz), 2.60(2H,m), 2.01(2H,m), 1.55(mH),1.20(mH), 0.85(mH), 0.10(6H,2s).

Compound 87

R_(f) :0.08 [hexanes:ethyl acetate, 1:1 (v/v)]

¹ H NMR (CDCl₃) δ: 7.51 ppm (1H,d,J=9.52 Hz), 5.95(4H,m), 5.30(mH),4.95(1H,m), 4.82(2H,m), 4.55(mH), 4.35(1H,m), 4.25(2H,m), 3.85(mH),3.65(mH), 3.16(mH), 2.84(1H, d,J=14.41 Hz), 2.55(mH), 2.01(mH),1.55(mH), 1.20(mH), 0.85(6H,t).

Compound 90

R_(f) :0.39 [methylene chloride:methyl alcohol, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ: 7.62 ppm (1H,d,J=8.0 Hz), 7.20(1H,d,J=8.1 Hz),5.90(7H,m), 5.46(1H,t,J=10.0 Hz), 5.40-5.15(mH), 5.05(1H,m), 4.85(mH),4.50(mH), 4.25(mH), 4.15(mH), 3.90(mH), 3.70(1H,m), 3.60(1H,m),3.53(1H,dd), 3.35(mH), 2.70-2.42(8H,m), 2.25(2H,t), 2.00(mH),1.80-1.45(mH), 1.37-1.07(mH), 0.85(15H,m).

Analog B235

R_(f) :0.47 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.43 min.

¹ H NMR (CDCl₃ :CD₃ OD, 1:1, v/v) δ: 5.15 ppm (2H, m), 5.00-4.80(3H,m),3.94-3.45(mH), 3.20-3.00(mH), 2.40-1.95(10H,m), 1.31-0.75(mH),0.55(15H,br.s).

¹³ C NMR (CDCl₃ :CD₃ OD, 1:1, v/v) δ: 208.3 ppm, 207.8, 175.6, 174.1,172.4, 170.0, 169.4, 102.8, 95.2, 76.8, 74.8, 74.3, 72.6, 71.8, 70.0,61.5, 55.0, 52.2, 43.8, 43.5, 43.0, 40.2, 38.5, 35.7, 33.2, 30.8, 30.6,30.4, 30.2, 26.9, 26.5, 26.4, 26.3, 24.8, 23.9, 23.7, 15.0.

Analog B235 (fully protected)

R_(f) :0.39 [methylene chloride:methyl alcohol, 19:1 (v/v)]

¹ H NMR (CDCl₃) δ: 7.40 ppm (1H,d), 7.13(1H,d), 5.93(7H,m), 5.70(1H,m),5.46-5.16(mH), 5.00(1H,m), 4.86(2H,m), 4.67-4.46(mH), 4.38-4.27(mH),4.18(1H,m), 3.93(1H,d), 3.80-3.60(mH), 3.40-3.30(mH), 2.68-2.42(mH),2.22(2H,t), 1.80-0.80(mH).

Analog B272

R_(f) :0.48 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.13 min.

Analog B272 (fully protected)

R_(f) :0.66 [methylene chloride:methyl alcohol, 19:1 (v/v/v/v)]

¹ H NMR (CDCl₃) δ: 7.13 ppm (1H,d), 7.05(1H,d), 5.92(8H,m), 5.69(1H,dd),5.32(mH), 5.09(1H,m), 4.88(1H,d), 4.80(1H,t), 4.65-4.45(mH),4.38-4.20(mH), 3.88-3.62(mH), 3.32(2H,q), 2.58(mH), 2.43(2H,m),2.25(2H,t), 2.00(4H,m), 1.58(mH), 1.23(mH), 0.85(15H,m).

Analog B286

R_(f) :0.43 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.70 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 6.58 ppm (2H,m), 5.61(2H, d, J=15.3Hz), 5.28(1H,dd,J=2.9,5.7 Hz), 5.20-5.07(2H,m), 5.00-4.88(3H,m),4.52(1H,d,J=8.2 Hz), 4.20-4.05(mH), 3.85-3.20(mH), 2.48-1.75(mH),1.40(mH), 1.20-1.00(mH), 0.67(15H,m).

³¹ P NMR (CDCl₃ :CD₃ OD 3:1, v/v) δ: 1.25 ppm, -1.61.

Analog B286 (fully protected)

R_(f) :0.60 [methylene chloride:methyl alcohol, 95:5 (v/v)]}

¹ H NMR (CDCl₃) δ: 6.88-6.73 ppm (4H,m), 5.98-5.85(7H,m), 5.83(1H,d),5.71(2H,m), 5.42-5.20(16H,m), 5.16(1H,t), 4.98(1H,m), 4.85(2H,dd),4.67-4.45(mH), 4.41-4.29(2H,m), 4.18(1H,m), 3.95(1H,dd), 3.88(1H,d),3.75(1H,m), 3.67(1H,m), 2.63-2.46(3H,m), 2.22(2H,t), 2.12(2H,t),2.00(2H,m), 1.78-1.15(mH), 0.84(15H,m).

Analog B287

R_(f) :0.49 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT (HPLC):13.70 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, (v/v) δ: 5.28 ppm (1H,dd,J=3.6,6.0 Hz),5.12(2H,m), 4.96(2H,m), 4.87(1H,m), 4.50(1H,d,J=8.7 Hz), 4.05(mH),3.80(mH), 3.60-3.24(mH), 2.40-2.10(mH), 1.80(mH), 0.65(15H,m).

³¹ P NMR (CDCl₃ :CD₃ OD 3:1, (v/v) δ: 0.31 ppm, -1.66.

Analog B287 (fully protected)

R_(f) :0.68 [methylene chloride:methyl alcohol, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ: 7.14 ppm (1H,d,J=7.9 Hz), 6.80(1H,d,J=8.0 Hz),5.92(8H,m), 5.70(1H,m), 5.45-5.17(mH), 5.05(1H,m), 5.02(1H,m),4.93(1H,d,J=7.9 Hz), 4.86(1H,t), 4.67-4.47(mH), 4.32(mH), 4.19(1H,dd),3.87(1H,d), 3.66(mH), 3.32(2H,q), 2.70-2.24(mH), 2.00(mH), 1.55(mH),1.25(mH), 0.85(15H,m).

Analog B288

R_(f) :0.82 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.48 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.30 ppm (1H,br.s),5.25-5.10(2H,m), 4.99(1H,t,J=9.5 Hz), 4.74(1H,t,J=9.4 Hz),4.39(1H,d,J=8.3 Hz), 4.02(1H,m), 3.82(2H,m), 3.71-3.10(mH),2.50-2.10(mH), 1.90-1.70(6H,m), 1.50-0.90(mH), 0.70(15H,t).

¹³ C NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 205.13 ppm, 205.02, 173.56,172.44, 170.78, 167.76, 167.32, 130.64, 130.54, 100.68, 94.57, 75.75,74.65, 72.58, 72.26, 70.10, 68.01, 67.83, 67.55, 60.74, 53.24, 51.04,42.87, 41.62, 38.46, 38.31, 36.64, 33.74, 33.60, 33.33, 33.18, 32.02,31.71, 31.63, 31.37, 31.32, 31.27, 31.20, 31.04, 30.86, 29.15, 29.11,29.08, 29.00, 28.97, 28.93, 28.88, 28.83, 28.80, 28.70, 28.59, 28.51,28.43, 28.30, 26.67, 25.00, 24.59, 24.37, 22.85, 22.78, 22.11, 21.97,13.37. ³¹ P NMR (CDCl₃ :CD₃ OD 3:1, v/v) δ: -1.49 ppm.

Analog B294

R_(f) :0.69 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

Analog B294 (fully protected)

R_(f) :0.40 [methylene chloride:methyl alcohol, 95:5 (v/v)]

RT (HPLC):15.07 min.

¹ H NMR (CDCl₃) δ: 7.41 ppm (1H,d,J=8.2 Hz), 7.09(1H,d,J=8.5 Hz),6.00-5.82(7H,m), 5.70(1H,m), 5.43-5.20(mH), 5.02(1H,m), 4.87(2H,m),4.69-4.44(mH), 4.40-4.28(3H,m), 4.18(1H,dd), 3.94(1H,d),3.78-3.61(4H,m), 3.46(2H,m), 3.38-3.28(5H,m), 2.65-2.42(mH), 2.00(5H,m),1.64-1.25(mH), 0.86(15H,m).

Analog B300

R_(f) :0.51 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.65 min.

Analog B300 (fully protected)

R_(f) :0.36 [methylene chloride:methyl alcohol, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ: 7.31 ppm (1H,d,J=8.0 Hz), 6.50(1H,d,J=8.1 Hz),6.00-5.83(7H,m), 5.78(1H,m), 5.46-5.20(14H,m), 5.00-4.84(3H,m),4.64-4.46(mH), 4.39-4.28(2H,m), 4.13(1H,m), 3.99(1H,d), 3.90(1H,m),3.75-3.60(3H,m), 3.47(1H,m), 3.38-3.30(2H,m), 2.65-2.43(6H,m),2.30(1H,dd), 2.18(1H,dd), 2.00(2H,t,) 1.60-1.20(mH), 0.87(15H,m).

Analog B313

R_(f) :0.57 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):11.75 min.

Analog B313 (fully protected)

R_(f) :0.71 [methylene chloride:methyl alcohol, 19:1 (v/v)]

¹ H NMR (CDCl₃) δ: 7.37 ppm (1H,d), 6.82(2H,m), 5.91(7H,m), 5.75(1H,t),5.7(1H,d), 5.41-5.19(mH), 4.96-4.87(2H,m), 4.63-4.48(mH), 4.45-4.30(mH),4.18(1H,m), 3.95(1H,d), 3.80-3.60(mH), 3.38(2H,q), 2.65-2.48(6H,m),2.24(2H,t), 2.17-1.95(6H,m), 1.70-1.10(mH), 0.85(15H,m).

Analog B314

R_(f) :0.48 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):27.93 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.26 ppm (1H,m), 5.17(1H,m),5.08(1H,m), 5.00(1H,m), 4.93(2H,m), 4.48(1H,d,J=7.9 Hz), 4.00(mH),3.85-3.20(mH), 1.80(4H,m), 1.40-1.00(mH), 0.70(18H,m).

Analog B314 (fully protected)

R_(f) :0.46 [methylene chloride:methyl alcohol, 19:1 (v/v)]

¹ H NMR (CDCl₃) δ: 7.38 ppm (1H,d,J=7.8 Hz), 6.14(1H,d,J=8.0 Hz),6.05-5.82(7H,m), 5.70(1H,t), 5.45-5.20(mH), 5.11-4.98(1H,m),4.90-4.82(2H,m), 4.67-4.42(mH), 4.39-4.22(mH), 4.10(1H,dd),3.98-3.85(2H,m), 3.83-3.62(mH), 3.38(2H,q), 2.68-2.30(mH), 2.25(mH),2.00(4H,m), 1.55(mH), 1.25(mH), 0.85(15H,m).

Analog B318

R_(f) :0.48 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.05 min.

Analog B318 (fully protected)

R_(f) :0.44 [methylene chloride:methyl alcohol, 19:1 (v/v)]

¹ H NMR (CDCl₃) δ: 7.40 ppm (1H,d), 6.99(1H,d), 6.05-5.82(7H,m),5.78(1H,q), 5.46-5.15(mH), 5.02(1H,m), 4.91-4.80(3H,m), 4.70-4.42(mH),4.38-4.20(mH), 3.90-3.25(mH), 2.93-2.50(mH), 2.25(2H,t), 2.00(mH),1.90-1.10(mH), 0.85(15H,m).

Analog B377

R_(f) :0.52 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

Analog B377 (fully protected)

R_(f) :0.39 [methylene chloride:methyl alcohol, 19:1 (v/v)]

¹ H NMR (CDCl₃) δ: 7.62 ppm (1H,d,J=8.0 Hz), 7.20(1H,d,J=8.1 Hz),5.90(7H,m), 5.46(1H,t,J=10.0 Hz), 5.40-5.15(mH), 5.05(1H,m), 4.85(mH),4.50(mH), 4.30(mH), 4.25(mH), 4.15(mH), 3.90(mH), 3.70(1H,m),3.60(1H,m), 3.53(1H,dd), 3.35(mH), 2.70-2.42(8H,m), 2.25(2H,t),2.00(mH), 1.80-1.45(mH), 1.37-1.07(mH), 0.85(15H,m).

Analog B379

R_(f) :0.55 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT (HPLC):9.12 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:2, v/v) δ: 5.14 ppm (2H,m), 5.05(1H,m),4.92(1H,t), 4.78(2H,m), 4.43(2H,m), 3.92-3.44(mH), 3.13(2H,m),2.93(1H,m), 2.70(1H,m), 2.35(3H,m), 2.22(4H,m), 2.12(1H,dd), 1.94(2H,m),1.74(2H,m), 1.30-0.90(mH), 0.58(15H,t).

Analog B379 (fully protected)

R_(f) :0.28 [methylene chloride:methyl alcohol, 100:3 (v/v)]

¹ H NMR (CDCl₃) δ: 7.62 ppm (1H,d), 7.08(1H,d), 5.90(7H,m), 5.70(1H,dd),5.40-5.20(17H,m), 5.00(1H,m), 4.82(2H,m), 4.65-4.46(15H,m), 4.35(2H,m),4.20(1H,dd), 3.90(1H,m), 3.67(1H,m), 3.35(3H,m), 3.20(1H,m), 3.10(1H,m),2.70(1H,dd), 2.58(2H,m), 2.47(3H,m), 2.26(2H,m), 2.03(2H,m),1.65-1.10(mH), 0.85(15H,t).

Analog B385

R_(f) :0.58 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):18.55 min.

Analog B385 (fully protected)

R_(f) :0.40 [methylene chloride:methyl alcohol, 97:3 (v/v)]

¹ H NMR (CDCl₃) δ: 6.52 ppm (1H,d), 5.93-5.85(7H,m), 5.71(1H,m),5.59(1H,br.s), 5.47(1H,br.s), 5.45-5.21(mH), 5.18(1H,m), 4.99(1H,m),4.83(2H,m), 4.66-4.45(mH), 4.40-4.25(mH), 4.18(1H,m), 3.95-3.65(mH),2.70-2.46(mH), 2.25(2H,t), 2.15-1.99(mH), 1.67-1.20(mH), 0.85(15H,m).

Analog B387

R_(f) :0.50 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):19.05 min.

Analog B387 (fully protected)

R_(f) :0.75 [methylene chloride:methyl alcohol, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ: 6.60 ppm (1H,d), 5.99-5.86(7H,m), 5.70(1H,m),5.44(1H,m), 5.42-5.21(mH), 5.12(1H,d), 4.65-4.44(mH), 4.35-4.26(mH),4.05(mH), 3.90(mH), 2.72-2.48(mH), 2.35(mH), 2.25(mH), 2.14-1.94(mH),1.58-1.18(mH), 0.89(15H,m).

Analog B388

R_(f) :0.60 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.72 min.

Analog B388 (fully protected)

R_(f) :0.80 [methylene chloride:methyl alcohol, 19:1 (v/v)]

¹ H NMR (CDCl₃) δ: 7.09 ppm (1H,d), 6.84(2H,m), 6.00-5.86(7H,m),5.84(1H,dd,J=3.0,17.0 Hz), 5.69(1H,dd,J=3.9,5.2 Hz), 5.46-5.22(mH),5.18(1H,dd), 5.01(1H,m), 4.86(1H,d,J=7.4 Hz), 4.80(1H,t,J=9.4 Hz),4.68-4.46(mH), 4.40-4.25(2H,m), 4.20(1H,m), 3.99(1H,q), 3.87(1H,d,J=11.5Hz), 3.75(1H,dd), 3.68(1H,m), 3.58(2H,dd), 3.34(2H,d), 2.69-2.50(4H,m),2.45(2H,t), 2.23(2H,t), 2.12(2H,m), 2.08-1.98(4H,m), 1.67-1.16(mH),0.88(15H,m).

Analog B398

R_(f) :0.49 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):7.12 min.

¹ H NMR (CD₃ OD) δ: 5.42 ppm (1H,m), 5.37(1H,t), 5.38(1H,m), 5.26(1H,m),5.18(1H,m), 4.70(1H,d,J=8.6 Hz), 4.22-3.82(mH), 3.76(1H,d,J=11.4 Hz),3.55(1H,t,J=11.0 Hz), 3.42(1H,d,J=10.3 Hz), 2.72-2.60(5H,m),2.42(1H,dd,J=8.6,17.1 Hz), 2.31(2H,t), 2.18-2.00(6H,m), 1.77-1.58(4H,m),1.50-1.23(mH), 0.87(15H,m).

¹³ C NMR (CD₃ OD) δ: 207.2 ppm, 206.8, 176.0, 174.7, 173.0, 170.8,170.2, 133.0, 130.8, 103.6, 96.2, 82.5, 82.4, 81.0, 80.8, 78.0, 77.9,77.8, 75.7, 75.6, 75.5, 74.8, 74.7, 73.3, 72.4, 72.1, 70.4, 70.2, 69.9,62.4, 56.8, 54.2, 44.5, 44.2, 44.0, 41.0, 39.7, 36.4, 35.6, 33.8, 33.5,32.0, 31.7, 31.2, 30.8, 29.0, 28.5, 27.8, 27.4, 27.3, 25.3, 25.1, 24.8,20.3, 20.0, 19.9, 15.5.

Analog B400

R_(f) :0.36 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.27 min.

Analog B400 (fully protected)

R_(f) :0.21 [methylene chloride:methyl alcohol, 98:2 (v/v)]

¹ H NMR (CDCl₃) δ: 7.43 ppm (1H,d), 7.08(1H,d), 6.98(1H,ddd),6.92(7H,m), 6.80(1H,dd), 6.70(1H,dd), 5.43-5.18(17H,m), 5.00(1H,m),4.87(2H,m), 4.65-4.30(mH), 4.18(1H,ddd), 3.93(1H,dd), 3.82(1H,q),3.70(2H,m), 3.33(4H,m), 2.58(2H,m), 2.46(4H,q), 2.17(2H,q),1.64-1.20(mH), 0.85(12H,t).

Analog B406

R_(f) :0.35 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.95 min.

Analog B406 (fully protected)

R_(f) :0.33 [methylene chloride:methyl alcohol, 19:1 (v/v)]

¹ H NMR (CDCl₃) δ: 7.32 ppm (1H,d), 7.17(1H,t), 6.05-5.85(7H,m),5.73(1H,m), 5.45-5.20(mH), 5.22(1H,m), 4.91-4.8(2H,m), 4.68-4.43(mH),4.40-4.28(mH), 4.20(1H,m), 3.91(1H,dd), 3.82-3.75(mH), 3.59(1H,q),3.42-3.29(mH), 2.72(2H,d), 2.35-2.20(mH), 2.25(2H,t), 2.10-1.91(mH),1.65(2H,t), 1.50(mH), 1.25(mH), 0.85(15H,m).

Analog B410

R_(f) :0.51 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.70 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.28 ppm (1H,br.s), 5.17(1H,m),5.09(1H,m), 5.00(1H,m), 4.95(1H,t,J=9.6 Hz), 4.46 (1H,d,J=8.1 Hz),4.09(1H,m), 3.85-3.46(mH), 3.25(3H,m), 2.45-2.25(6H,m), 2.06(2H,t),1.80(4H,m), 0.65(15H,m).

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 1.32 ppm, -1.12.

Analog B410 (fully protected)

R_(f) :0.41 [methylene chloride:methyl alcohol, 97:3 (v/v)]

¹ H NMR (CDCl₃) δ: 7.50 ppm (2H,dd), 5.92(7H,m), 5.80(1H,m),5.42-5.23(17H,m), 4.80(2H,m), 4.74(1H,t), 4.62-5.00(15H,m), 4.33(2H,m),4.11(2H,m), 3.85(1H,t), 3.68(2H,m), 3.42(2H,d), 3.36(2H,s), 2.78(2H,d),2.50(4H,q), 2.22(2H,t), 2.00(4H,m), 1.8-1.50(1H,m), 1.25(mH),0.85(15H,m).

Analog B415

R_(f) :0.50 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):12.62 min.

Analog B415 (fully protected)

R_(f) :0.20 [methylene chloride:methyl alcohol, 98:2 (v/v)]

¹ H NMR (CDCl₃) δ: 7.53 ppm (1H,d), 7.04(1H,d), 5.94(7H,m), 5.73(1H,m),5.43-5.20(17H,m), 5.02(1H,m), 4.88(2H,m), 4.58(15H,m) 4.37(1H,m),4.28(1H,dd), 4.18(1H,t), 4.12(1H,t), 3.95(1H,dd), 3.86(1H,t),3.78(1H,dd), 3.67(1H,m), 3.59(2H,m), 3.44(3H,m), 3.32(2H,q), 2.55(4H,m),2.45(2H,t), 2.02(4H,m), 1.78(2H,m), 1.68-1.20(mH), 0.87(15H,m).

Analog B425

R_(f) :0.62 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.05 min.

¹ H NMR (CD₃ OD:CDCl₃, 2:1, v/v) δ: 5.40-5.12 ppm (mH), 4.18-3.70(mH),3.45-3.19(mH), 2.60(mH), 2.25(2H,t), 2.00(mH), 1.80(mH), 1.65-1.15(mH),0.85(15H,m).

Analog B425 (fully protected)

R_(f) :0.75 [methylene chloride:methyl alcohol, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ: 7.45 ppm (1H,d), 6.84(1H,d), 5.95(7H,m), 5.80(1H,m),5.46-5.22(mH), 5.05(1H,m), 4.85(mH), 4.67-4.48(mH), 4.30(mH),3.95-3.80(mH), 3.75(2H,d), 3.65(1H,m), 3.25(mH), 3.15(2H,t), 2.65(mH),2.60(1H,d), 2.55(1H,d), 2.25(2H,t), 2.00(mH), 1.80(mH), 1.65-1.20(mH),0.85(15H,m).

Analog B426

R_(f) :0.44 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.63 min.

Analog B426 (fully protected)

R_(f) :0.50 [methylene chloride:methyl alcohol, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ: 7.38 ppm (1H,d), 7.11(1H,d), 5.95(7H,m), 5.72(1H,m),5.65(mH), 5.42-5.18(mH), 5.05(mH), 4.95(1H,t), 4.85(1H,d),4.68-4.25(mH), 3.95(1H,m), 3.79(mH), 3.55-3.30(mH), 2.68(2H,t),2.58(2H,t), 2.45(mH), 2.25(2H,t), 2.00(mH), 1.69-1.45(mH),1.30-1.15(mH), 0.85(15H,m).

Analog B427

R_(f) :0.62 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.17 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.37 ppm (1H,m), 5.27(1H,m),5.17(1H,m), 4.78(1H,m), 4.53(1H,d,J=6.3 Hz), 4.04-3.20(mH), 2.43(4H,m),2.15(2H,t), 1.95-1.83(4H,m), 1.70-0.95(mH), 0.75(15H,t).

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 1.24 ppm, -1.40.

Analog B442

¹ H NMR (CDCl₃ :CD₃ OD, 2:1, v/v) δ: 5.20 ppm (mH), 5.13(mH), 4.95(mH),4.75(mH), 3.85-3.28(mH), 2.68(mH), 2.40(mH), 2.10(2H,t), 1.80(mH),1.65-1.00(mH), 0.70(15H,m).

Analog B442 (fully protected)

¹ H NMR (CDCl₃) δ: 7.60-7.50 ppm (2H,2d), 5.95(7H,m), 4.88(1H,m),4.70-4.45(mH), 4.35(2H,m), 4.15(mH), 3.85-3.45(mH), 2.88(mH),2.65(2H,t), 2.25(2H,t), 2.00(mH), 1.78-1.23(mH), 0.85(15H,m).

Analog B451

R_(f) :0.45 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):12.37 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, (v/v) δ: 5.35 ppm (1H,dd,J=3.4,6.6 Hz),5.17(1H,m), 5.09(1H,m), 4.92(1H,t,J=10.3 Hz), 4.90(1H,m), 4.56(1H,J=8.4Hz), 4.00(2H,m), 3.80-3.20(mH), 3.18(3H,s), 3.17(1H,t), 2.68(4H,m),2.36(2H,ABX,J=4.5,8.4,16.1 Hz), 2.10(2H,t), 1.80(4H,m), 1.55-1.05(mH),0.67(15H,m).

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: -0.51 ppm, -1.41.

Analog B451 (fully protected)

R_(f) :0.19 [methylene chloride:methyl alcohol, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ: 7.74 ppm (1H, d), 7.54(1H,d), 5.92(6H,m), 5.85(1H,m),5.44-5.20(16H,m), 4.86(2H,m), 4.70(1H,t), 4.63(9H,m), 4.50(4H,m),4.38(1H,q), 4.17(1H,m), 4.08(1H,m), 3.82-3.46(8H,m), 3.38(3H,s),2.96-2.73(4H,m), 2.62(2H,m), 2.26(2H,m), 2.00(4H,m), 1.80-1.18(mH),0.85(15H,m).

Analog B452

R_(f) :0.44 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):10.30 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.34 ppm (1H,m), 5.18(1H,m),4.95(1H,t,J=9.6 Hz), 4.93(1H,m), 4.50(1H,d,J=8.3 Hz), 4.05(1H,m),3.92-3.60(mH), 3.50-3.20(mH), 2.65(2H,m), 2.40-2.10(mH), 1.85(4H,m),1.55-1.00(mH), 0.70(15H,m).

¹³ C NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 173.52 ppm, 172.22, 170.24,164.80, 164.62, 130.71, 127.87, 100.08, 94.05, 74.49, 73.09, 72.26,71.68, 69.72, 68.01, 67.49, 66.94, 59.99, 57.58, 56.96, 55.94, 53.83,51.80, 42.15, 38.61, 36.90, 34.07, 33.43. 32.03, 31.49, 31.40, 29.25,29.20, 28.93, 28.83, 28.58, 28.37, 28.27, 26.81, 26.12, 25.20, 24.70,24.54, 23.68, 22.24, 21.37, 13.52.

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 1.24 ppm, -1.65.

Analog B452 (fully protected)

R_(f) :0.47 [methylene chloride:methyl alcohol, 95:5, (v/v)]

¹ H NMR (CDCl₃) δ: 7.48 ppm (1H,d), 7.03(1H,d), 5.95(7H,m), 5.79(1H,m),5.45-5.18(mH), 5.03(1H,m), 4.85(mH), 4.75-4.50(mH), 4.30(mH),4.25(1H,m), 3.91(1H,dd), 3.78-3.60(mH), 3.55(mH), 3.30(2H,d),2.90-2.50(mH), 2.25(2H,t), 2.00(mH), 1.80-1.15(mH), 0.85(15H,m).

Analog B459

R_(f) 0.49 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.37-14.13 min (multiple peaks)

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.42 ppm(1H,br.s), 5.25(1H,m),5.17(1H,m), 4.75(1H,m), 4.64(1H,d,J=7.7 Hz), 4.53(1H,m), 4.00(1H,m),3.90-3.20(mH), 2.80(4H,m), 2.15(2H,t), 1.90-1.80(4H,m), 1.70-1.00(mH),0.75(15H,br.s).

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 1.39 ppm, -1.51.

Analog B459 (fully protected)

R_(f) :0.43 [methylene chloride, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ: 7.68-7.50 ppm (2H,m), 5.95(7H,m), 5.85(1H,m),5.42-5.23(12H,m), 4.93(1H,m), 4.82(1H,m), 4.76(1H,m), 4.66-4.55(mH),4.31(1H,m), 4.26(1H,q), 4.12(2H,m), 3.83-3.42(mH), 2.95-2.84(4H,m),2.26(2H,t), 2.00(4H,m), 1.80-1.18(mH), 0.85(15H,m).

Analog B460

R_(f) :0.63 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.52 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.34 ppm (1H,m), 5.19(1H,m),5.11(1H,m), 4.68(1H,m), 4.41(1H,d,J=8.1 Hz), 3.90(1H,m), 3.81-3.12(mH),2.70(4H,q), 2.10(2H,t), 1.80(4H,m), 1.58-0.90(mH), 0.65(15H,t).

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 1.38 ppm, -1.30

Analog B460 (fully protected)

¹ H NMR (CDCl₃) δ: 7.68 ppm (1H,d), 7.53(1H,d), 5.95(7H,m), 5.83(1H,m),5.45-5.21(mH), 4.95(1H,m), 4.82(mH), 4.72(2H,q), 4.55(mH), 4.28(mH),4.10(mH), 3.80(1H,d), 3.70-3.55(mH), 3.51-3.45(mH), 2.95-2.81(mH),2.25(2H,t), 2.00(mH), 1.75(mH), 1.45(mH), 1.25(mH), 0.85(15H,m).

Analog B465

R_(f) :0.83 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.53 min.

¹ H NMR (CDCl₃ :CD₃ OD, 2:1, v/v) δ: 5.34 ppm (1H,dd,J=3.3,6.7 Hz),5.19(1H,m), 5.11(1H,m), 4.69(1H,m), 4.54(1H,d,J=7.9 Hz), 3.92(1H,m),3.83-3.24(mH), 2.70(4H,m), 2.10(2H,t), 1.80(4H,m), 1.60-0.95(mH),0.65(15H,m).

³¹ P NMR (CDCl₃ :CD₃ OD, 2:1, v/v) δ: 1.32 ppm, -1.40.

Analog B465 (fully protected)

¹ H NMR (CDCl₃) δ: 7.55 ppm (2H,m), 5.93(7H,m), 5.80(1H,m),5.50-5.25(mH), 4.90(2H,m), 4.75-4.50(mH), 4.30(2H,m), 4.13(1H,m),3.85-3.40(mH), 2.95-2.80(4H,m), 2.26(2H,t), 2.00(4H,m), 1.80-0.80(mH).

Analog B466

R_(f) :0.51 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.45 min

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.36 ppm (1H,dd,J=3.2,6.7 Hz),5.18(1H,m), 5.09(1H,m), 4.65(1H,m), 4.48(1H,d,J=8.3 Hz), 3.90-3.24(mH),3.17(3H,s), 2.70(4H,q), 2.10(2H,t), 1.8- (4H,m), 1.55-1.00(mH),0.65(15H,t).

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: -0.67 ppm, -1.50.

Analog B466 (fully protected)

¹ H NMR (CDCl₃) δ: 7.67 ppm (1H,d), 7.55(1H,d), 6.03-5.88(6H,m),5.84(1H,m), 5.46-5.21(mH), 4.92(1H,m), 4.88-4.50(mH), 4.32(1H,q),4.10(1H,m), 3.88-3.43(mH), 3.37(3H,s), 3.0-2.79(2H,m), 2.30(2H,t),2.10-1.25(mH), 0.85(15H,m).

Analog B477

R_(f) :0.53 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20v/v/v/v)]

RT(HPLC):13.48 min

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.36 ppm (1H,dd,J=3.3,6.8 Hz),5.18(1H,m), 5.10(1H,m), 4.70(1H,m), 4.57(1H,d,J=8.2 Hz), 3.90-3.25(mH),3.20(3H,s), 2.73(4H,m), 2.10(2H,t), 1.80(4H,m), 1.65-0.90(mH),0.70(15H,t).

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: -0.64 ppm, -1.44.

Analog B477 (fully protected)

R_(f) :0.41 [methylene chloride:methyl alcohol, 95:5 (v/v)]

¹ H NMR (CDCl₃) δ: 7.56 ppm (2H,m), 5.93(6H,m), 5.82(1H,m),5.44-5.24(12H,m), 4.90(1H,m), 4.70(1H,t), 4.66-4.53(mH), 4.32(1H,q),4.12(1H,m), 3.85-3.42(mH), 3.38(3H,s), 2.93-2.82(mH), 2.26(2H,t),2.00(4H,m), 1.80-1.20(mH), 0.85(15H,m).

Analog B479

R_(f) :0.97 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

Analog B510

R_(f) :0.47 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):6.37 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.36 ppm (1H,br.s), 4.55(1H,d,J=8.2Hz), 4.00-3.20(mH), 3.23(3H,s), 2.40(4H,br.s), 1.60-0.70(mH).

¹³ C NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 205.96 ppm, 205.80, 167.85,167.19, 100.14, 94.74, 80.13, 78.70, 74.03, 73.61, 73.18, 70.50, 69.56,69.16, 68.85, 67.08, 58.21, 54.45, 52.20, 52.13, 42.89, 42.76, 37.26,37.23, 36.62, 36.36, 31.36, 30.42, 29.21, 29.09, 29.01, 28.95, 28.78,28.57, 28.51, 25.20, 25.10, 22.79, 22.09, 13.25.

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: -0.72 ppm, -1.49.

Analog B464

R_(f) :0.51 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):17.62 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.29 ppm (1H,m), 5.18(1H,m),5.09(1H,m), 4.70(1H,m), 4.44(mH), 3.88(2H,m), 3.72-3.12(mH), 3.16(3H,s),2.36(4H,m), 2.07(2H,t), 1.87-1.76(4H,m), 1.60-0.92(mH), 0.65(15H,m).

¹³ C NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 206.07 ppm, 206.00, 174.15,167.99, 167.15, 130.69, 127.82, 100.22, 94.98, 80.12, 78.86, 74.88,74.82, 73.72, 72.84, 71.81, 70.70, 69.74, 69.17, 67.28, 58.38, 55.12,54.21, 52.12, 43.08, 42.98, 42.78, 37.37, 36.65, 34.47, 34.19, 33.63,32.14, 31.48, 31.35, 29.41, 29.25, 29.22, 29.14, 29.08, 29.05, 28.93,28.91, 28.87, 28.72, 28.62, 28.54, 26.78, 26.13, 25.32, 24.82, 24.63,22.94, 22.88, 22.22, 13.47.

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: -0.629 ppm, -1.431.

Analog B587

R_(f) :0.62 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):14.80 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.26 ppm (1H,m), 5.17(1H,m),5.10(1H,m), 4.70(1H,m), 4.49(1H,d,J=8.10 Hz), 3.91-3.29(mH), 3.26(3H,s),3.24-3.20(mH), 3.17(3H,s), 3.12(mH), 2.89(1H,t,J=9.30 Hz), 2.34(4H,m),2.08(2H,t), 1.89-1.75(4H,m), 1.62-0.92(mH), 0.64(15H,m).

¹³ C NMR (CDCl₃ :CD₃ OD), 3:1, v/v) δ: 206.41 ppm, 205.56, 174.17,167.80, 167.18, 130.69, 127.83, 99.93, 94.62, 80.18, 79.50, 78.92,74.78, 73.84, 72.31, 71.83, 70.77, 69.84, 69.06, 68.27, 67.88, 66.93,64.48, 61.86, 59.96, 58.49, 54.98, 52.53, 50.60, 43.15, 42.89, 37.30,36.99, 34.44, 34.17, 33.64, 31.48, 31.40, 31.35, 29.34, 29.22, 29.13,29.08, 28.93, 28.84, 28.70, 28.61, 28.54, 26.79, 26.14, 25.29, 24.81,24.62, 22.92, 22.86, 22.22, 13.49.

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: -0.673 ppm, -1.509.

Analog B718

R_(f) :0.40 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.65 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1 v/v) δ: 5.34 ppm (1H,br.s), 5.20(1H,m),5.12(1H,m), 4.70(2H,m), 4.32(1H,d,J=8.0 Hz), 4.0-3.85(mH),3.70-3.20(mH), 3.18(3H,s), 3.22-3.13(mH), 2.35(mH), 2.08(2H,t),1.90(3H,s), 1.90-1.77(4H,m), 1.63-1.00(mH), 0.67(mH,t).

¹³ C NMR (CDCl₃ :CD₃ OD, 3:1 v/v) δ: 206.35 ppm, 205.88, 174,23, 169.94,168.25, 167.20, 130.80, 127.94, 100.45, 94.44, 80.48, 74.68, 74.38,73.93, 71.89, 70.65, 69.70, 69.28, 69.20, 68.39, 67.02, 58.59, 54.85,52.19, 43.23, 43.07, 37.31, 36.97, 34.50, 34.26, 33.74, 31.50, 31.45,31.08, 29.43, 29.37, 29.32, 29.28, 29.20, 29.17, 29.14, 29.04, 29.01,28.94, 28.84, 28.71, 28.64, 26.89, 26.25, 25.37, 24.89, 24.72, 23.03,22.97, 22.33, 22.30, 20.43, 13.78, 11.98.

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: -0.63 ppm, -1.59.

Analog B725

R_(f) :0.58 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):17.58 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.32 ppm (1H,br.s), 5.24(1H,m),5.12(1H,m), 4.78(1H,m), 4.36(1H,d), 4.04-3.00(mH), 3.34(3H,s),2.36(4H,m), 2.10(2H,t), 1.85(4H,m), 1.60-1.10(mH), 0.67(15H,t).

¹³ C NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 206.06 ppm, 205.94, 174.02,167.82, 167.20, 130.87, 127.82, 101.11, 94.88, 81.24, 79.45, 78.93,77.30, 73.69, 72.58, 71.53, 70.04, 69.82, 69.40, 69.01, 68.02, 64.72,59.95, 55.31, 52.30, 49.00, 48.78, 48.57, 48.36, 48.14, 47.93, 47.72,43.15, 43.07, 37.39, 36.67, 34,89, 34.21, 33.75, 31.54, 31.45, 31.42,29.41, 29.27, 29.20, 29.15, 28.99, 28.88, 28.77, 28.70, 28.61, 26.87,26.22, 25.37, 24.89, 24.74, 23.03, 22.96, 22.29, 22.26, 13.57.

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 0.74 ppm, -1.27.

Analog B736

R_(f) :0.57 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):12.57 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.20 ppm (1H,m), 5.10(1H,m),4.70(1H,m), 4.53(1H,d,J=3.5 Hz), 4.40(1H,d,J=7.7 Hz), 3.90-3.20(mH),3.18(3H,s), 2.34(4H,q), 2.10(2H,t), 1.85(4H,m), 1.70-0.90(mH),0.65(15H,m).

¹³ C NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 206.00 ppm, 205.81, 174.05,167.52, 167.01, 130.62, 127.71, 100.47, 97.33, 80.00, 79.66, 74.71,73.69, 71.67, 70.67, 70.56, 69.64, 69.42, 69.24, 69.12, 67.84, 66.51,65.00, 58.35, 58.34, 55.21, 52.02, 43.05, 42.88, 37.23, 36.60, 34.42,34.10, 33.55, 31.39, 31.37, 31.30, 31.25, 29.26, 29.17, 29.12, 29.08,29.03, 28.99, 28.95, 28.86, 28.84, 28.81, 28.74, 28.63, 28.55, 28.45,26.70, 26.04, 25.23, 24.74, 22.88, 22.80, 22.13, 22.10, 13.38.

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 0.99 ppm, -0.48.

Analog B737

R_(f) :0.71 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):12.45 min.

¹ H NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 5.31 ppm (1H,m), 5.29(1H,m),5.10(1H,m), 4.71(1H,m), 4.46(1H,d,J=8.0 Hz), 4.20(1H,t),4.07(1H,t,J=4.14 Hz), 3.98-3.81(mH), 3.72-3.27(mH), 3.17(3H,s),3.12(mH), 2.33(4H,m), 2.07(2H,t), 1.90-1.78(mH), 1.60-0.98(mH),0.65(15H,m).

¹³ C NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 206.1 ppm, 179.7, 174.1, 168.0,167.2, 130.7, 127.8, 100.3, 94.2, 90.3, 84.0, 80.1, 74.8, 73.8, 71.8,70.6, 70.4, 70.1, 69.5, 69.1, 68.1, 66.9, 66.7, 64.7, 58.2, 55.1, 51.9,43.0, 42.9, 37.3, 36.7, 34.4, 34.2, 33.6, 32.5, 31.5, 31.4, 31.3, 29.3,29.2, 29.1, 29.0, 28.9, 28.8, 28.7, 28.6, 28.5, 28.4, 26.8, 26.1, 25.3,24.8, 24.6, 22.9, 22.2, 13.5.

³¹ P NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: -0.635 ppm, -1.634.

¹⁹ F NMR (CDCl₃ :CD₃ OD, 3:1, v/v) δ: 1.62 ppm.

Analog B763

R_(f) :0.92 [chloroform:methyl alcohol:acetic acid:water, 125:75:10:20(v/v/v/v)]

RT(HPLC):13.70 min.

Compound 92

R_(f) :0.26 [hexanes:ethyl acetate, 4:1 (v/v)]

¹ H NMR (CDCl₃) δ: 5.95 ppm (1H,m), 5.32(1H,d,J=17.2 Hz),5.28(1H,d,J=10.4 Hz), 4.80(mH), 4.58(2H,d), 4.50(1H,d,J=7.40 Hz),4.08(1H,q), 3.78-3.49(mH), 3.30-3.13(mH), 2.30(1H_(OH),s), 2.06(3H,s),1.80(mH), 1.55(mH), 1.25(mH), 0.89(9H,s), 0.83(3H,t), 0.11(6H,s).

Compound 93

¹ H NMR (CDCl₃) δ: 5.92 ppm (1H,m), 5.36(1H,dd,J=1.4, 17.3 Hz),5.28(1H,dd,J=1.2, 10.5 Hz), 4.88(1H,m), 4.62(2H,m), 4.48(1H,d,J=7.6 Hz),3.87-3.67(mH), 3.50(3H,s), 3.22-3.13(3H,m), 3.07(1H,dd,J=8.3, 9.7 Hz),1.90(3H,m), 1.60(2H,m), 1.23(mH), 0.90(9H,s), 0.85(3H,t), 0.12(6H,2s).

Compound 94

R_(f) :0.69 [hexanes:ethyl acetate, 2:1 (v/v)]

¹ H NMR (CDCl₃) δ: 5.94(1H,m), 5.35(1H,d,J=15.5 Hz), 5.25(1H, d,J=9.3Hz), 4.89(1H,m), 4.62(2H,m), 4.54(1H,d,J=7.4 Hz),4.46-4.28(1H,ddd,J=7.7,10.1,50.0 Hz), 3.90-3.68(mH), 3.43(1H,m),3.33-3.19(2H,m), 1.88(2H,m), 1.77(1H,t), 1.26(mH, br.s), 0.88(9H,s),0.86(3H,t), 0.15(6H,2s).

Compound 95

R_(f) :0.06 [hexanes:ethyl acetate, 1:1 (v/v)]

¹ H NMR (CDCl₃) δ: 5.93 ppm (4H,m), 5.27(8H,m), 4.92(1H,d,J=3.5 Hz),4.78(1H,m), 4.70-4.55(8H,m), 4.21(1H,m), 3.88-3.60(mH),3.59(1H,dd,J=4.6,13.0 Hz), 3.30(1H,dd,J=3.5,10.1 Hz), 1.83(2H,m),1.54(2H,m), 1.26(mH), 0.85(3H,m).

Compound 96

R_(f)α :0.52 [hexanes:ethyl acetate, 1:1 (v/v)] R_(f)β :0.30[hexanes:ethyl acetate, 1:1 (v/v)]

¹ H NMR (CDCl₃) δ: 8.69 ppm (1H,s), 6.33(1H.sub.α,d,J=3.4 Hz),5.92(2H,m), 5.56(1H.sub.β,d,J=8.5 Hz), 5.30(6H,m), 5.02(1H,m),4.53(4H,m), 4.31(1H,m), 4.22(2H,m), 3.82(3H,m), 3.71(1H,t,J=8.5 Hz),3.57(3H.sub.α,s), 3.56(3H.sub..sub.β,s), 3.49(2H,m), 3.25(3H,m),2.10(2H,m), 2.04(4H,m), 1.88(2H,m), 1.65(2H,m), 1.56(1H,s), 1.25(mH),0.86(6H,m).

Compound 97

¹ H NMR (CDCl₃) δ: 8.74 ppm (1H,s), 6.40(1H₁α,d), 5.59(1H.sub.β,d,J=8.5Hz), 5.40-5.23(6H,m), 5.0(1H,m), 4.57(4H,m), 4.50(1H₄α,q), 4.36(1H₄β,q),3.80(3H,m), 3.63(3H,m), 3.42(2H,m), 3.30(1H,t,J=9.3Hz), 2.28(2H,t),2.08-1.82(mH), 1.70-1.20(mH), 0.83(9H,m).

Compound A1

¹ H NMR (CDCl₃) δ: 4.96 ppm (1H_(enone-H),s), 4.19(2H,q), 3.74(2H,s),3.43(2H,s), 2.52(2H,t), 2.17(2H_(enone) form,t), 1.68-1.51(2H,m),1.37-1.18(9H,m), 0.88(3H,t).

Compound A2

¹ H NMR (CDCl₃) δ: 4.16 ppm (2H,q), 3.98(1H,m), 2.95(1H,d),2.52(1H,dd,J=2.9,5.8 Hz), 2.49(2H,dd), 1.55-1.51(2H,m), 1.47-1.37(2H,m),1.30-1.11(10H,m), 0.87(3H,t).

Compound A3

MP:105.6°-106.2° C.

¹ H NMR (CDCl₃) δ: 3.86-3.78 ppm (1H,m), 2.96-2.86(2H,m), 2.33,2.16(2H,ABX,J=2.4,9.5,15.6 Hz), 2.03-1.94(4H,br.d), 1.83-1.74(4H,br.d),1.66-1.60(2H,br.d), 1.54-1.10(22H,m), 0.86(3H,t).

Compound A4

¹ H NMR (CDCl₃) δ: 7.92 ppm (2H,dd,J=1.1,7.7 Hz), 7.63(1H,td,J_(t) =7.6Hz,J_(d) =1.2 Hz), 7.50(2H, J=7.7 Hz), 5.49, 5.38(2H,AB,J=16.5 Hz),4.14(1H,m), 3.50(1H,br.s), 2.69-2.58(2H,ABX,J=2.9,9.4,15.1 Hz),1.65-1.45(2H,m), 1.4-1.2(10H,m), 0.88(3H,t).

Compound A5

¹ H NMR (CDCl₃) δ: 7.91 ppm (2H,dd,J=1.2,6.4 Hz), 7.61(1H,td,J_(t) =7.6Hz,J_(d) =1.2 Hz), 7.50(2H,t,J=7.8 Hz), 5.99-5.89(1H,m),5.42,5.38(2H,AB,J=15.1 Hz), 5.27(2H,d), 5.24(1H,m), 4.6(2H,dd),2.83,2.76(2H,ABX,J=5.37,7.56,21.7 Hz), 1.75-1.65(2H,m),1.41-1.26(10H,m), 0.87(3H,t).

Compound A6

¹ H NMR (CDCl₃ :CD₃ OD, 15:1) δ: 5.90 ppm (1H,m), 5.32(2H,dd),5.19-5.01(1H,m), 4.57(2H,dt), 2.61-2.56(2H,ABX,J=5.37,7.57,19.7 Hz),1.65-1.55(2H,m), 1.31-1.21(10H,m), 0.82(3H,t).

Compound A7

¹ H NMR (CDCl₃) δ: 4.90 ppm (1H_(enone-H),s), 3.74(3H,s), 3.45(2H,s),2.52(2H,t), 2.18(2H_(enone) form,t), 1.62-1.52(2H,m), 1.35-1.20(8H,m),0.88(3H,t).

Compound A8

¹ H NMR (CDCl₃) δ: 4.08 ppm (1H,m), 3.71(3H,s), 2.88(1H,d,J=3.8 Hz),2.49(1H,dd,J=3.1,16.4 Hz), 2.41(1H,dd,J=9.1,16.5 Hz), 1.58-1.47(2H,m),1.44-1.38(2H,m), 1.37-1.23(10H,m), 0.87(3H,t).

Compound A9

¹ H NMR (CDCl₃) δ: 3.91-3.75 ppm (3H,m), 2.62-2.38(2H_(OH),m),1.75-1.61(2H,m), 1.55-1.36(2H,m), 1.35-1.23(10H,m), 0.87(3H,t).

Compound A10

¹ H NMR (CDCl₃) δ: 7.86 ppm (2H,d,J=8.3 Hz), 7.34(2H,d,J=8.1 Hz),4.27(1H,m), 4.13(1H,m), 3.72(1H,m), 2.44(3H,s), 1.89-1.81(1H,m),1.68-1.62(2H,m), 1.39-1.25(12H,m), 0.87(3H,t).

¹³ C NMR (CDCl₃) δ: 144.44 ppm, 132.59, 129.49, 127.49, 67.60, 37.14,35.86, 31.39, 29.11, 28.83, 25.11, 22.25, 21.26, 13.70.

Compound A11

¹ H NMR (CDCl₃) δ: 3.21 ppm (2H,t), 2.20-2.10(4H,m), 1.93(2H,m),1.58(2H,m), 1.46(2H,m), 1.40-1.22(6H,m), 0.88(3H,t).

Compound A12

¹ H NMR (CDCl₃) δ: 7.43 ppm (2H,d), 7.33-7.20(10H,m), 6.84(2H,d),3.79(3H,s), 3.74(1H,br.s), 3.38(1H,m), 3.22(1H,m), 2.98(1H,d,J=2.9 Hz),1.72(1H,m), 1.56-1.24(10H,m), 0.87(3H,t).

Compound A13

¹ H NMR (CDCl₃) δ: 7.44 ppm (2H,d), 7.33-7.19(10H,m), 6.82(2H,d),3.79(3H,s), 3.40(2H,m), 3.28(1H,m), 3.15(2H,t), 2.11(4H,q), 1.75(2H,q),1.55-1.25(29H,m), 0.88(6H,t).

Compound A14

¹ H NMR (CDCl₃) δ: 3.80 ppm (2H,m), 3.52(2H,m), 3.42(1H,m), 2.72(1H,m),2.17(4H,m), 1.80-1.25(26H,m), 0.88(6H,t).

Compound A16

¹ H NMR (CDCl₃) δ: 3.68 ppm (1H,m), 3.48(2H,t), 2.52(2H,m), 2.14(4H,m),1.68-1.26(24H,m), 0.87(6H,t).

Compound A17

¹ H NMR (CDCl₃) δ: 5.33 ppm (2H,m), 3.68(1H,m), 3.52(2H,m), 2.56(2H,m),2.02(2H,m), 1.98-1.27(27H,m), 0.88(6H,t).

Compound A18

¹ H NMR (CDCl₃) δ: 7.80 ppm (2H,d), 7.34(2H,d), 4.05(2H,t), 2.51(2H,m),2.45(3H,s), 2.06(2H,m), 1.41(2H,m), 1.27(6H,m), 0.88(3H,t).

Compound A19

¹ H NMR (CDCl₃) δ: 7.84 ppm (2H,m), 7.72(2H,m), 3.83(2H,t), 2.56(2H,m),2.06(2H,m), 1.34(2H,m), 1.20(6H,m), 0.84(3H,t).

Compound A20

¹ H NMR (CDCl₃) δ: 7.82 ppm (2H,m), 7.71(2H,m), 5.44(1H,m), 5.37(1H,m),3.72(2H,t), 2.44(2H,q), 1.95(2H,m), 1.18(8H,m), 0.83(3H,t).

Compound A21

¹ H NMR (CDCl₃) δ: 5.48 ppm (1H,m), 5.34(1H,m), 2.71(2H,t), 2.18(2H,q),2.03(2H,m), 1.27(8H,m), 0.87(3H,t).

Compound A22

¹ H NMR (CDCl₃) δ: 7.91 ppm (2H,d), 7.61(1H,t,J=7.3 Hz), 7.56(2H,t),5.50(1H,m), 5.35(2H,s), 5.30(1H,m), 5.15(1H,t), 4.78(1H,t), 3.17(2H,m),2.74(2H,t), 2.24(2H,t), 2.01(2H,q), 1.58(2H,m), 1.57(2H,d), 1.26(16H,m),0.87(6H,t).

Compound A23

¹ H NMR (CD₃ OD) δ: 6.85 ppm (1H,m), 5.45(1H,m), 5.32(1H,m), 5.06(2H,m),4.92(1H,d), 3.15-3.00(mH), 2.45(2H,t), 2.32(1H,d), 2.20(mH), 2.03(mH),1.59(mH), 1.28(mH), 0.85(6H,m).

Compound A24

¹ H NMR (CDCl₃) δ: 7.90 ppm (2H,d), 7.61(1H,t), 7.49(2H,t), 5.34(2H,s),5.30(1H,m), 2.74(2H,m), 2.31(2H,t), 1.69-1.57(4H,m), 1.37-1.20(28H,m),0.88(6H,t).

Compound A25

¹ H NMR (CDCl₃) δ: 5.21 ppm (1H,m), 2.62(2H,m), 2.29(2H,t), 1.61(4H,m),1.36-1.18(26H,m), 0.89(6H,t).

Compound A26

¹ H NMR (CDCl₃) δ: 3.76 ppm (3H,s), 2.32(2H,t), 1.56(2H,m),1.40-1.35(mH), 1.30-1.22(mH), 0.88(3H,t).

Compound A27

¹ H NMR (CDCl₃) δ: 5.65 ppm (1H,s), 3.67(3H,s), 2.61(2H,t), 1.88(3H,s),1.50-1.40(mH), 1.35-1.20(mH), 0.88(3H,t).

Compound A28

¹ H NMR (CDCl₃) δ: 5.66 ppm (1H,s), 3.68(3H,s), 2.15(2H,t),1.50-1.40(mH), 1.34-1.20(mH), 0.88(3H,t).

Compound A29

¹ H NMR (CDCl₃) δ: 5.40 ppm (1H,t,J=7.1 Hz), 4.15(2H,t), 2.00(2H,t),1.66(3H,s), 1.40-1.20(18H,m), 0.88(3H,t).

Compound A30

¹ H NMR (CD₃ OD) δ: 5.64 ppm (1H,s), 4.91(1H,br.s), 2.15(2H,t),2.11(3H,s), 1.48(4H,m), 1.29(14H,m), 0.89(3H,t).

Compound A31

R_(f) :0.76 [hexanes:ethyl acetate, 3:2 (v/v)]

¹ H NMR (CDCl₃) δ: 6.02 ppm (1H,s), 2.19(2H,m), 2.13(3H,s), 1.48(2H,m),1.26(16H,m), 0.88(3H,t).

Compound B1

¹ H NMR (CDCl₃) δ: 3.30 ppm (2H,m), 2.26(2H,m), 2.13(1H,m), 1.95(1H,m),1.47(mH), 1.28(mH), 0.88(3H,t).

Compound B2

¹ H NMR (CDCl₃) δ: 2.50 ppm (2H,t), 2.33(2H,m), 2.13(2H,m), 1.83(2H,m),1.45(2H,m), 1.30-1.25(6H,m), 0.89(3H,t).

Compound B3

¹ H NMR (CDCl₃) δ: 2.49 ppm (2H,t), 2.24(2H,m), 2.13(2H,m), 1.80(2H,m),1.45(2H,m), 1.37-1.26(6H,m), 0.88(3H,t).

Compound B4

¹ H NMR (CDCl₃) δ: 5.42 ppm (1H,m), 5.32(1H,m), 2.37(2H,t), 2.10(2H,m),2.01(2H,m), 1.70(2H,m), 1.28(8H,m), 0.89(3H,t).

Compound B5

¹ H NMR (CDCl₃) δ: 7.90 ppm (2H,d), 7.61(1H,t), 7.49(2H,m), 5.38(1H,m),5.30(2H,m), 2.75(2H,m), 2.31(2H,m), 2.06(2H,m), 1.99(2H,m), 1.68(2H,m),1.27(mH), 0.88(6H,t).

Compound B6

¹ H NMR (CDCl₃) δ: 5.38 ppm (1H,m), 5.21(1H,m), 2.61(2H,m), 2.29(2H,t),2.05(2H,m), 1.99(2H,m), 1.67(2H,m), 1.62(2H,m), 1.26(15H,m), 0.87(6H,t).

Compound C1

¹ H NMR (CDCl₃) δ: 4.19 ppm (2H,q), 3.43(2H,s), 2.52(2H,m), 1.60(3H,m),1.29(18H,m), 0.87(3H,t).

Compound C2

R_(f) :0.35 [hexanes:ethyl acetate, 4:1 (v/v)]

¹ H NMR (CDCl₃) δ: 4.16 ppm (2H,q), 3.97(1H,m), 2.46(1H,dd,J=3.2,16.4Hz), 2.38(1H,dd,J=9.0,16.4 Hz), 1.54-1.10(23H,m), 0.86(3H,t).

Compound C3

¹ H NMR (CDCl₃) δ: 3.84 ppm (1H,m), 2.96(2H,m), 2.36(1H,dd,J=2.7,15.9Hz), 2.17(1H,dd,J=9.3,15.6 Hz), 2.02(4H,m), 1.78(4H,m), 1.66(2H,m),1.41(mH), 1.25(mH), 0.87(3H,t).

Compound C4

¹ H NMR (CDCl₃) δ: 7.93 ppm (2H,d), 7.63(1H,t,J=7.3 Hz), 7.50(2H,t),5.43(2H,q), 4.14(1H,m), 2.70(1H,dd,J=2.9,14.9 Hz), 2.53(1H,dd,J=9.3,15.1Hz), 1.40-1.20(20H,m), 0.88(3H, t).

Compound C5

¹ H NMR (CDCl₃) δ: 7.91 ppm (2H,2d), 7.59(1H,t,J=6.5 Hz), 7.49(2H,t),7.27(2H,d), 6.86(2H,d), 5.33(2H,q), 4.51(2H,q), 3.94(1H,m), 3.79(3H,s),2.79(1H,dd,J=7.0,15.2 Hz), 2.66(1H, dd,J=5.5,15.3 Hz), 1.62(2H,m),1.40-1.26(16H,m), 0.88(3H,t).

Compound C6

¹ H NMR (CDCl₃) δ: 6.87 ppm (4H,2d), 4.50(2H,s), 3.85(1H,p,J=5.9 Hz),3.79(3H,s), 2.60-2.55(2H,m), 1.69-1.60(1H,m), 1.59-1.59(1H,m),1.40-1.18(18H,m), 0.88(3H,t).

Compound C7

¹ H NMR (CDCl₃) δ: 4.15 ppm (2H,q), 3.05(4H,m), 2.70(2H,m) 2.04(3H,m),1.87(1H,m), 1.31-1.21(21H,m), 0.87(3H,t).

Compound C8

¹ H NMR (CDCl₃) δ: 3.12 ppm (2H,s), 3.03(2H,t), 2.76(2H,t),2.13-2.01(mH), 1.89(mH), 1.54(mH), 1.26(mH), 0.88(3H,t).

Compound D1

¹ H NMR (CDCl₃) δ: 7.36 ppm (5H,s), 5.17(2H,s), 3.48(2H,s), 2.50(2H,t),1.56(6H,s), 1.24(12H,m), 0.88(3H,t).

Compound D2

¹ H NMR (CDCl₃) δ: 3.52 ppm (2H,s), 2.56(2H,t), 1.16(2H,m), 1.25(14H,m),0.88(3H,t).

Compound E1

¹ H NMR (CDCl₃) δ: 3.74 ppm (3H,s), 3.22(2H,s), 2.62(2H,t), 1.59(2H,m),1.35(2H,m), 1.25(14H,m), 0.88(3H,t).

Compound E2

¹ H NMR (CDCl₃) δ: 3.80 ppm (3H,s), 3.68(2H,m), 2.82(2H,m), 1.77(2H,m),1.50-1.40(2H,m), 1.26(14H,m), 0.88(3H,t).

Compound E3

¹ H NMR (CDCl₃) δ: 3.84 ppm (1H,d,J=14.6 Hz), 3.49(1H,d,J=14.4 Hz),3.08-3.03(1H,m), 2.89-2.81(1H,m), 1.79-1.74(2H,m), 1.51-1.40(2H,m),1.35-1.25(14H,m), 0.88(3H,t).

Compound E4

¹ H NMR (CDCl₃) δ: 3.80 ppm (3H,s), 3.68(2H,m), 2.84(2H,m), 1.77(2H,m),1.50-1.40(2H,m), 1.26(14H,m), 0.88(3H,t).

Compound E5

¹ H NMR (CDCl₃) δ: 3.82-3.68 ppm (2H,m), 3.07-2.86(2H,m), 1.75(2H,m),1.45(2H,m), 1.29(mH), 0.87(3H,t).

Compound E6

¹ H NMR (CDCl₃) δ: 3.96 ppm (2H,s), 3.82(3H,s), 3.24(2H,m), 1.86(2H,m),1.44(mH), 1.25(mH), 0.87(3H,t).

Compound E7

R_(f) :0.33 [methylene chloride:methyl alcohol, 19:1 (v/v)]

¹ H NMR (CDCl₃) δ: 4.01 ppm (2H,s), 3.27(2H,m), 1.87(2H,m),1.47-1.26(16H,m), 0.88(3H,t).

Compound G1

¹ H NMR (CDCl₃) δ: 4.19 ppm (2H,q), 3.45(2H,s), 2.67(2H,t), 2.20(2H,m),2.12(2H,m), 1.76(2H,m), 1.45(2H,m), 1.35-1.25(9H,m), 0.88(3H,t).

Compound G2

¹ H NMR (CDCl₃) δ: 4.21 ppm (2H,m), 3.06(2H,m), 2.94(2H,q),2.20-2.01(mH), 1.80-1.50(mH), 1.48-1.43(mH), 1.39-1.22(mH), 0.88(3H,t).

EXAMPLE 3 In Vitro Inhibition of LPS-Induced Production of TumorNecrosis Factor (TNF) and IL-1β

Both bacterial LPS and bacterial lipid A elicit production of tumornecrosis factor (TNF) and IL-1β in cultured human monocytes (J. Immunol.139:429, 1987). The lipid A analogs described herein inhibit such LPS-and/or lipid A-mediated induction as demonstrated by the followingexperiments.

Monocytes were isolated from human blood by Percoll density gradientcentrifugation, plated at approximately 1×10⁶ cells/well on a 48-wellplate in RPMI 1640 medium (GIBCO, Grand Island, N.Y.) containing 10%human serum (Sigma Chemical Co., St. Louis, Mo.) and incubated for twoto three hours. Bacterial LPS (i.e., from E. coli 0111:B4; SigmaChemicals, St. Louis, Mo.) at 10 ng/ml or lipid A (Daiichi Chemicals,Tokyo, Japan) at 1.0 ng/ml in RPMI 1640 medium were combined with 0.45ml of RPMI 1640 medium containing 1% human serum and added to thecultured monocytes. In experiments involving a lipid A analog, theanalog was added immediately before addition of LPS or lipid A invarying concentrations (e.g., between 0 and 100 μM in a 50 μl aliquot).Following a three-hour incubation period, a 0.1 ml aliquot of theculture supernatant was assayed for the presence of TNF and IL-1β. TNFand IL-1β were assayed using the ELISA assay of R & D Systems(Minneapolis, Minn.) and the instructions of the manufacturer, however,any other standard ELISA kits may be utilized, for example, the kitavailable from Genzyme, Cambridge, Mass. Experiments were performed intriplicate.

The lipid A analogs inhibited LPS-induced production of TNF in humanmonocytes in a concentration dependent manner. Of the lipid A analogstested, Lipid A Analog B531-35 was found to be one of the most effectivecompounds to inhibit LPS-induced production of TNF, exhibiting an ED₅₀of approximately 0.02 nM. Other lipid A analogs found to inhibitLPS-induced TNF production included B214-32, B410-32, B442-32, B451-32,B452-32, B427-32, B459-32, B460-32, B464-32, B464-34, B464-35, B465-32,B466-32, B477-32, B477-35, B718-35, B587-35, B737-35, B736-35, B725-35,and B763-35; these compounds exhibited ED₅₀ s of between 0.03 nM and 129nM.

Lipid A analogs similarly inhibited the LPS-induced production of IL-1βin human monocytes. LPS was added at 10 ng/ml, and lipid A analogs wereadded at a concentration of between 0 and 10 μM. Inhibition of IL-1βproduction was also found to be concentration dependent.

In a separate set of experiments, LPS-induced TNF production wasinhibited by lipid A analogs in macrophages isolated from guinea pigsand mice. Hartley-White guinea pig (Elm Hill Breeders, Chelmsford,Mass.) and C57BL/6 mouse (Jackson Labs, Bar Harbor, Me.) macrophageswere isolated from the abdomen of primed animals. Priming wasaccomplished by intraperitoneal injection of 2 mg of Bacillus calmetteguerin (BCG; RIBI Immunochemical Research, Inc., Hamilton, Mont.) at aconcentration of 10 mg/ml in physiological saline for mice and 2 mg ofBCG at a concentration of 2 mg/7 ml in mineral oil for guinea pigs.Three days post-injection, peritoneal macrophages were isolated from theabdomen of the animals by standard techniques. Cells were allowed toadhere to culture plates for two to three hours and were then contactedwith RPMI 1640 medium containing 10% fetal calf serum and LPS (at 10ng/ml). To test inhibition, lipid A analogs (at a concentration ofbetween 0 and 100 μM) were added to the culture medium just prior to LPSaddition. Following a three-hour incubation period, guinea pig and mouseTNF levels were assayed by the cytolytic bioassay described inLymphokines 2:235, 1981. Lipid A analogs B214-32, B410-32, B442-32,B451-32, B452-32, B427-32, B459-32, B460-32, B464-32, B464-34, B464-35,B465-32, B466-32, B477-32, B477-35, and B718-35 (all analogs tested todate) similarly inhibited LPS-induced TNF production in both guinea pigsand mice. Analogs B464-34 and B531-35 provided the most effectiveinhibition in guinea pigs (ED₅₀ 's=0.04 nM and 0.66 nM, respectively);analogs B477-32 and B531-35 provided very effective inhibition in mice(ED₅₀ 's=1.3 nM and and 2.26 nM, respectively). Lipid A analogs B214-32,B410-32, B442-32, B451-32, B452-32, B427-32, B459-32, B460-32, B464-32,B464-34, B464-35, B465-32, B466-32, B477-32, B477-35, and B718-35 (allanalogs tested to date) inhibited LPS-induced TNF production. ED₅₀ 'smeasured in the experiments involving guinea pig macrophages ranged fromapproximately 0.04 nM to 18.5 nM. ED₅₀ 's measured in the experimentsinvolving mouse macrophages ranged from approximately 1.0 nM to 1.0 μM.

EXAMPLE 4 In Vivo Assays

BCG-primed mice (as described above) were utilized as an in vivo assaysystem for monitoring the inhibitory effects of lipid A analogs on (1)LPS-induced TNF production and (2) LPS-induced lethality as follows.

Five week old male C57BL/6 mice (supra) were primed by intravenous tailvein injection with 2 mg of BCG. Ten days post-injection, E. coli LPS(supra) in pyrogen-free 5% glucose solution (Otsuka PharmaceuticalsInc., Tokyo, Japan) was administered intravenously through the tail veinof the BCG-primed mice. LPS was administered at a concentration of 1-3μg/mouse for both TNF production and mortality studies. In experimentsinvolving a lipid A analog, the analog was administered as a componentof the injected LPS solution at a concentration of between 10 and 300μg/mouse. Plasma was obtained one hour post-LPS injection, and TNF wasassayed by the ELISA assay described above. Mortality resulting fromseptic shock was recorded for 36 hours post-LPS injection.

Lipid A analogs effectively suppressed the production of TNF followingadministration of LPS. Analogs B318-32 and B531-35 effectively inhibitedTNF production in vivo in mice (ED₅₀ 's=5.4 μg/mouse and 16.2 μg/mouse,respectively). Analogs B214-31, B214-32, B214-33, B218-32, B231-32,B235-32, B262-32, B274-32, B278-32, B286-32, B294-32, B313-32, B314-32,B318-32, B379-32, B380-32, B398-32, B399-32, B400-32, B406-32, B410-32,B415-32, B425-32, and B426-32 also inhibited TNF production.

In parallel experiments carried out in guinea pigs, these analogs werealso effective inhibitors of LPS-induced TNF production in vivo (optimumED₅₀ 's=7.5 μg/guinea pig and 5 μg/guinea pig measured for analogB214-32).

EXAMPLE 5 Lipid A Analogs Suppress LPS-Stimulated Virus Production

LPS potently stimulates the production of viruses which reside (in theirlatent phase) in monocytes or macrophages (see, e.g., Pomerantz et al.,J. Exp. Med. 172:253, 1990; Masihi et al., J. of Acquired Imm.Deficiency Syndromes 3:200, 1990). In the case of HIV-1, increased viralproduction likely results from activation of cells by both a directactivation by LPS and the LPS-mediated elevation in TNF-α levels.Cellular activation promotes increased binding of trans-acting factorsto the HIV-1 NF-κB binding site; this, in turn, leads to increased viraltranscription and replication (see, e.g., Duh et al., Proc Natl. Acad.Sci. USA 86:5974, 1989).

The lipid A analogs described herein inhibited an LPS-mediated increasein HIV-1 replication. This was demonstrated using an in vitro modelsystem which monitored HIV-1 long terminal repeat (LTR) transcription.Because activation of the transcriptional enhancer of the HIV-1 longterminal repeat (LTR) has been correlated with viral replication (Colmanet al., AIDS 2:185, 1988; Nabel and Baltimore, Nature 326:711, 1987),the assay provides a reliable measure of viral replication, and hencevirus production.

Plasmid HIV-1-LTR-CAT (Pomerantz et al., 1990, supra), a construct whichincludes the HIV-1 LTR fused, in frame, to the chloramphenicolacetyltransferase (CAT) gene, was propagated in HB-101 cells (Gibco-BRL,Grand Island, N.Y.). Plasmid was purified from host cell extract using aQiagen affinity column and the instructions of the manufacturer (QiagenInc., Chatsworth, Calif.) and transiently transfected into U937 cells(ATCC Accession No. CRL 1593; American Type Culture Collection;Rockville, Md.) generally by the method of Pomerantz et al. (1990,supra), except that 80 μM chloroquine (Promega Biotech, Madison, Wis.)was present throughout the transfection procedure, and 10⁶ U937 cellswere transfected with either 20 μg of HIV-1-LTR-CAT or 10 μg of pCAT(i.e., a control plasmid carrying only the CAT gene; Promega Biotech,Madison, Wis.). Twenty-four hours post-transfection, cells wereincubated with or without a lipid A analog (typically at a concentrationof between of 0.0 and 1.0 μM). Following a 30 minute incubation phorbolmyristate acetate (PMA; Sigma Chemical Co., St. Louis, Mo.) at 50 ng/mland E. coli 0111:B4 LPS (described above) at 100 ng/ml were added to thecells, and incubation was allowed to proceed for an additional 24 hours.Cells were then harvested and lysed as described in Ausubel et al.(Current protocols in Molecular Biology, John Wiley & Sons, New York,N.Y., 1987), extract protein concentration was determined using theMicro BCA Protein Assay System of Pierce Chemical Co. (Rockville, Ill.),and CAT activity assayed as the rate of acetylation of chloramphenicol(in DPM/min) as described in Biotech Update 5(2):28 (Novel FluorDiffusion CAT Assay Facilitates Sample Processing and Analysis, DuPontCo. Publication, Biotechnology Systems, Wilmington, Del.). Lipid Aanalog B398-32 inhibited the LPS-mediated induction of LTR-CATtranscription with an average IC₅₀ of 85 nM.

Similar results were obtained using U937 cell lines stably transfectedwith an HIV-1 LTR-CAT fusion gene. Experiments were carried out usingone such stably-transfected cell line, i.e., the U938 cell line ofLatham et al. (Cell. Immunol. 129:513, 1990). Cells were cultured asdescribed in Latham et al. (1990, supra), and 10⁶ cells were treatedwith a lipid A analog (at a concentration of between 0.0 and 1.0 μM).Thirty minutes after addition of the analog, cells were treated withphorbol myristate acetate (at a concentration of 0.33 ng/ml; describedabove) and E. coli 0111:B4 LPS (at a concentration of 33 ng/ml;described above). Cells were cultured for an additional 24 hours,harvested, and assayed for CAT activity as described above. Resultsindicate that B477 suppresses the LPS-stimulated activation of the HIVLTR at an IC₅₀ of 15 nM. Analogs B398-32, B400-32, B427-32, B464-32, andB466-32, similarly suppressed LPS-stimulated HIV LTR activation withIC₅₀ s ranging from 15 to 260 nM. B464-32 similarly inhibited activationof the HIV-LTR mediated by the LPS of other Gram-negative bacteria (forexample, Salmonella typhimurium).

NF-κB-regulated transcriptional control is not unique to HIV-1. Otherviral genomes, including that of Simian Virus-40 (SV-40), include anNF-κB binding site within an enhancer element of its early promoter(Nakamura et al., J. Biol. Chem. 264:20189, 1989). A plasmid constructcontaining an SV40 promoter enhancer-CAT fusion gene, termed pCAT(Promega Biotech), was transiently transfected into U937 cells asdescribed above. 10⁶ cells/plate were treated with a lipid A analog (ata concentration of 0.0, 0.1, or 1.0 μM). Thirty minutes after additionof the analog, cells were treated with phorbol myristate acetate (at aconcentration of 50 ng/ml) and E. coli 0111:B4 LPS (at a concentrationof 100 ng/ml). Cells were cultured for an additional 24 hours,harvested, and assayed for CAT activity as described above.LPS-stimulated CAT expression was inhibited or completely blocked bylipid A analog B398-32. In another "Experiment 4", LPS-stimulated CATexpression was inhibited or completely blocked by lipid A analogB466-32. These results indicated that the lipid A analogs describedherein effectively suppressed the LPS-mediated increase in SV-40replication.

The lipid A analogs described herein may similarly suppress theactivation of any virus whose replication is directly or indirectlycontrolled by an NF-κB regulatory region. Such viruses include, withoutlimitation, cytomegaloviruses or Herpes viruses (e.g., Herpes simplex).In addition, because influenza virus activation (in monocytes andmacrophages) is potentiated by LPS (Nain et al., J. Immunol. 145:1921,1990) and an enhanced release of TNF-α has been implicated in observedcomplications of combined influenza A and bacterial infections, theinstant lipid A analogs are likely to suppress influenza virusactivation as well.

THERAPY

The lipid A analogs described herein provide useful therapeutics for thetreatment or prevention of any LPS-mediated disorder. Such disordersinclude without limitation: endotoxemia (or sepsis syndrome) resultingfrom a Gram⁻ bacteremia (with its accompanying symptoms of fever,generalized inflammation, disseminated intravascular coagulation,hypotension, acute renal failure, acute respiratory distress syndrome,hepatocellular destruction, and/or cardiac failure); and LPS-mediatedexacerbation of latent or active viral infections (e.g., infection withHIV-1, cytomegaloviruses, herpes simplex viruses, and influenza virus).

The lipid A analog is typically administered in apharmaceutically-acceptable formulation, e.g., dissolved inphysiological saline or physiological saline which may include 5%glucose (for the purpose of increased analog solubility). Administrationis by any appropriate route, but, ordinarily, it will be administeredintravenously, either by intravenous injection or transfusion. When thelipid A analog is provided for the treatment of a viral infection, itmay be administered in conjunction with appropriate viricidal agents.The lipid A analogs may be stored as a freeze-dried formulation.

Lipid A analogs are administered in dosages which provide suitableinhibition of LPS activation of target cells; generally, these dosagesare, preferably, between 0.001-500 mg/patient, more preferably, between0.01-300 mg/patient and, most preferably, between 0.01-100 mg/patient.

What is claimed is:
 1. A compound of formula: ##STR290## wherein R² is:##STR291## wherein each J, independently, is OH or a protected OH; eachL is O, N, or C; each M is O or N; each E, independently, is an integerbetween 0 and 14 inclusive; each m, independently, is an integer between0 and 14 inclusive; each n, independently, is an integer between 0 and14 inclusive; each p, independently, is an integer between 0 and 10inclusive; each q, independently, is an integer between 0 and 10inclusive; each x, independently, is an integer between 0 and 14inclusive; each y, independently, is an integer between 0 and 14inclusive; each z, independently, is an integer between 0 and 10inclusive; and each G, independently, is N, O, S, SO, or SO₂ ;P¹ is OH,a protected OH, or a protected A¹ group wherein each A₁ group,independently, is: ##STR292## wherein each d, independently, is aninteger between 0 and 5 inclusive; each f, independently, is an integerbetween 0 and 5 inclusive; each g, independently, is an integer between0 and 5 inclusive; and each A³, independently, is: ##STR293##

    (CH.sub.2).sub.j --CO.sub.2 H, or O--(CH.sub.2).sub.j --CO.sub.2 H

wherein each j, independently, is an integer between 0 and 14 inclusive;and P² is H, a halo group, OH, a protected OH, O(CH₂)_(w) CH₃,##STR294## wherein w is an integer between 0 and 14 inclusive, or apharmaceutically acceptable salt thereof.
 2. The compound of claim 1,wherein R² is: ##STR295## wherein each J, independently, is OH or aprotected OH; each m, independently, is an integer between 0 and 10inclusive; each n, independently, is an integer between 0 and 10inclusive; each x, independently, is an integer between 0 and 10inclusive; each z, independently, is an integer between 0 and 3inclusive; each G, independently, is SO or SO₂ ; and for each p and q,independently,

    ≦ (p+q)≦12;

P¹ is OH, a protected OH, or a protected A¹ group wherein each A¹ groupis: ##STR296## wherein each d, independently, is an integer between 0and 2 inclusive; and P² is H, OH, a protected OH, or O(CH₂)_(w) CH₃,wherein w is an integer between 0 and 3 inclusive.
 3. The compound ofclaim 2, wherein R² is ##STR297## wherein each J, independently, is OHor a protected OH; each x, independently, is an integer between 6 and 11inclusive; each G, independently, is SO or SO₂ ; each n, independently,is an integer between 6 and 10 inclusive; and

    ≦ (p+q)≦10;

P¹ is ##STR298## and P² is H, OH, a protected OH, or OCH₃.
 4. A compoundof the formula: ##STR299## wherein R⁴ is ##STR300## wherein each J,independently, is OH or a protected OH; L is O, N, or C; each M is O orN; each E, independently, is an integer between 0 and 14 inclusive; eachm, independently, is an integer between 0 and 14 inclusive; each n,independently, is an integer between 0 and 14 inclusive; each p,independently, is an integer between 0 and 10 inclusive; each q,independently, is an integer between 0 and 10 inclusive; each x,independently, is an integer between 0 and 14 inclusive; each y,independently, is an integer between 0 and 14 inclusive; each z,independently, is an integer between 0 and 10 inclusive; and each G,independently, is N, O, S, SO, or SO₂ ;Z is OH, a protected OH, anactivated OH, or a displacable leaving group; P³ is OH, a protected OH,OCH₃, A^(2'), or a protected A^(2'), wherein said A^(2') group is:##STR301## wherein each d, independently, is an integer between 0 and 5inclusive; each f, independently, is an integer between 0 and 5inclusive; each g, independently, is an integer between 0 and 5inclusive; and each A³, independently, is: ##STR302##

    (CH.sub.2).sub.j --CO.sub.2 H, or O--(CH.sub.2).sub.j --CO.sub.2 H

wherein each j, independently, is an integer between 0 and 14 inclusive;X' is X or a protected X group, wherein said X group is H, (CH₂)_(t)CH₃, (CH₂)_(t) OH, (CH₂)_(t) O(CH₂)_(v) CH₃, (CH₂)_(t) OPO(OH)₂,

    (CH.sub.2).sub.t --CH═CH--(CH.sub.2).sub.v CH.sub.3, (CH.sub.2).sub.t --O--R.sup.5, ##STR303## wherein each t and v, independently, is an integer between 0 and 14 inclusive; and R.sup.5 is any of the possibilities listed above for R.sup.1 -R.sup.4, or a pharmaceutically acceptable salt thereof.


5. The compound of claim 4 wherein R⁴ is: ##STR304## wherein each J,independently, is OH or a protected OH; each m, independently, is aninteger between 0 and 10 inclusive; each n, independently, is an integerbetween 0 and 10 inclusive; each x, independently, is an integer between0 and 10 inclusive; each z, independently, is an integer between 0 and 3inclusive; each G, independently, is SO or SO₂ ; and for each p and q,independently,

    ≦ (p+q)≦12;

each P³ is H, OH, a protected OH, A^(2') or protected A^(2'), whereinA^(2') is ##STR305## wherein each d, independently, is an integerbetween 0 and 2 inclusive; and X' is H, (CH₂)_(t) OH, (CH₂)_(t)O(CH₂)_(v) CH₃ or (CH₂)_(t) CH₃, wherein t is an integer between 0 and 6inclusive and v is an integer between 0 and
 6. 6. The compound of claim5, wherein R⁴ is ##STR306## wherein each J, independently, is OH or aprotected OH; each x, independently, is an integer between 6 and 11inclusive; and each G, independently, is SO or SO₂ ; each n,independently, is an integer between 6 and 10 inclusive; and

    ≦ (p+q)≦10;

P³ is OH, a protected OH, A^(2'), or a protected A^(2'), wherein A^(2')is ##STR307## and X' is CH₂ OH, CH₂ OCH₃, or CH₂ O(CH₂)_(v) CH₃, whereinv is an integer between 1 and 3 inclusive.